Overview

Publication Information

The published version of this manuscript can be found here: Sieler Jr M.J., Al-Samarrie C.E., Kasschau K.D., Kent M.L., Sharpton T.J.. Modelling the zebrafish gut microbiome’s resistance and sensitivity to climate change and infection. bioRxiv (2025). DOI:

All code generated during this analysis is available in the GitHub Repository.

The raw sequence files generated during the current study are available at the NCBI Sequence Read Archive (SRA) project numbers: PRJNA1219243.

Fig. 1) Experimental Design Schematic

Fig.1 Experimental design showing treatments and husbandry events during the course of the study. Symbols indicate when an experimental event occurred at each time point. (1) 260 fish were assigned and acclimated to one of three water temperature groups (e.g., 28°C, 32°C, or 35°C) and reared from 0 to 164 days post fertilization (dpf). (2a) At 164 dpf (or 0 days post exposure; DPE), fecal collections were collected from a random selection of five fish per tank (n = 60). Additionally, histological and wet mount assessments were conducted on selected fish to assess presence of infection and infection burden. (1b) Afterwards, a cohort of fish from each water temperature group were exposed to Pseudocapillaria tomentosa. (3-6) Subsequent fecal samples were collected and histopathological assessments were conducted at 14 dpe (n = 54), (3) 21 dpe (n = 48), (4) 28 dpe (n = 47), and (5) 42 dpe (n = 51).

Results

Fig. 2) Water temperature shapes gut microbiome structure

Fig. 2 Effects of water temperature on zebrafish gut microbiomes. (A) Simpson’s Index of diversity shows that gut microbiome diversity significantly differs between fish reared at 28°C and 35°C water temperatures. (B) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main effect of temperature. The analysis shows that gut microbiome composition significantly differs between fish reared at different water temperatures. (C) Simpson’s Index of diversity shows microbial gut diversity increases with time from 0 days post exposure (dpe) to 42 dpe, irrespective of water temperature. (D) Capscale ordination of gut microbiome composition based on the Bray-Curtis dissimilarity constrained on the main effects of water temperature and time (days post exposure, dpe), and their interaction. The analysis shows that shows that gut microbiome composition differs between fish across time depending on water temperature. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate direction of greatest change in the indicated by covariates.

2A)

Tables
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GLM
GLM Results
glm(Alpha.Score ~ Temperature); Unexposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −0.754 0.091 −8.309 <0.001 ****
Temperature32 0.051 0.127 0.400 ≥0.25 ns
Temperature35 0.054 0.129 0.421 ≥0.25 ns
Simpson
(Intercept) −0.101 0.092 −1.103 ≥0.25 ns
Temperature32 0.132 0.128 1.030 ≥0.25 ns
Temperature35 0.340 0.131 2.591 0.010 *
Richness
(Intercept) −0.231 0.075 −3.079 0.002 **
Temperature32 0.047 0.105 0.446 ≥0.25 ns
Temperature35 −0.473 0.110 −4.299 <0.001 ****
Phylogenetic
(Intercept) −0.255 0.072 −3.540 <0.001 ***
Temperature32 0.068 0.101 0.673 ≥0.25 ns
Temperature35 −0.475 0.106 −4.482 <0.001 ****
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature), type = 2); Unexposed fish
term statistic df p.value sig
Shannon
Temperature 0.225 2.000 ≥0.25 ns
Simpson
Temperature 6.830 2.000 0.033 *
Richness
Temperature 27.512 2.000 <0.001 ****
Phylogenetic
Temperature 31.449 2.000 <0.001 ****
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature); Unexposed fish
term .y. group1 group2 estimate std.error statistic adj.p.value Variable
Shannon
Temperature Alpha.Score 32 28 0.051 0.127 0.400 ≥0.25 Temperature
Temperature Alpha.Score 35 28 0.054 0.129 0.421 ≥0.25 Temperature
Temperature Alpha.Score 35 32 0.004 0.127 0.029 ≥0.25 Temperature
Simpson
Temperature Alpha.Score 32 28 0.132 0.128 1.030 ≥0.25 Temperature
Temperature Alpha.Score 35 28 0.340 0.131 2.591 0.026 Temperature
Temperature Alpha.Score 35 32 0.207 0.130 1.596 0.247 Temperature
Richness
Temperature Alpha.Score 32 28 0.047 0.105 0.446 ≥0.25 Temperature
Temperature Alpha.Score 35 28 −0.473 0.110 −4.299 <0.001 Temperature
Temperature Alpha.Score 35 32 −0.520 0.109 −4.768 <0.001 Temperature
Phylogenetic
Temperature Alpha.Score 32 28 0.068 0.101 0.673 ≥0.25 Temperature
Temperature Alpha.Score 35 28 −0.475 0.106 −4.482 <0.001 Temperature
Temperature Alpha.Score 35 32 −0.543 0.105 −5.172 <0.001 Temperature
Addl. Metrics

2B)

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ADONIS2
ADONIS2
adonis2(Beta Distance ~ Temperature); Unexposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 2.294 0.133 12.900 0.001 ***
Residual 168.000 14.936 0.867 NA NA NA
Total 170.000 17.229 1.000 NA NA NA
canberra
Temperature 2.000 3.862 0.099 9.178 0.001 ***
Residual 168.000 35.344 0.901 NA NA NA
Total 170.000 39.206 1.000 NA NA NA
gunifrac
Temperature 2.000 1.422 0.102 9.566 0.001 ***
Residual 168.000 12.485 0.898 NA NA NA
Total 170.000 13.907 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Unexposed fish
term df sumsq meansq statistic p.value sig
bray
Temperature 2.000 0.008 0.004 0.386 ≥0.25 ns
Residual 168.000 1.771 0.011 NA NA NA
canberra
Temperature 2.000 0.008 0.004 0.873 ≥0.25 ns
Residual 168.000 0.759 0.005 NA NA NA
gunifrac
Temperature 2.000 0.005 0.003 0.369 ≥0.25 ns
Residual 168.000 1.143 0.007 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature); Unexposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temperature 32 28 −0.011 −0.056 0.034 ≥0.25 ns
Distance Temperature 35 28 0.005 −0.041 0.051 ≥0.25 ns
Distance Temperature 35 32 0.016 −0.029 0.062 ≥0.25 ns
canberra
Distance Temperature 32 28 −0.001 −0.031 0.028 ≥0.25 ns
Distance Temperature 35 28 0.014 −0.016 0.044 ≥0.25 ns
Distance Temperature 35 32 0.015 −0.015 0.045 ≥0.25 ns
gunifrac
Distance Temperature 32 28 −0.013 −0.049 0.023 ≥0.25 ns
Distance Temperature 35 28 −0.004 −0.040 0.033 ≥0.25 ns
Distance Temperature 35 32 0.009 −0.027 0.046 ≥0.25 ns
Addl. Metrics

2C)

Tables
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GLM
GLM Results
glm(Alpha.Score ~ Temperature*DPE); Unexposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −1.074 0.138 −7.754 <0.001 ****
Temperature32 0.075 0.193 0.386 ≥0.25 ns
Temperature35 0.154 0.193 0.797 ≥0.25 ns
DPE 0.018 0.006 3.086 0.002 **
Temperature32:DPE −0.001 0.008 −0.170 ≥0.25 ns
Temperature35:DPE −0.005 0.008 −0.585 ≥0.25 ns
Simpson
(Intercept) −0.433 0.134 −3.245 0.001 **
Temperature32 0.157 0.186 0.841 ≥0.25 ns
Temperature35 0.408 0.187 2.177 0.031 *
DPE 0.019 0.006 3.293 0.001 **
Temperature32:DPE −0.001 0.008 −0.173 ≥0.25 ns
Temperature35:DPE −0.003 0.008 −0.343 ≥0.25 ns
Richness
(Intercept) −0.651 0.108 −6.017 <0.001 ****
Temperature32 0.189 0.150 1.257 0.211 ns
Temperature35 −0.059 0.154 −0.383 ≥0.25 ns
DPE 0.024 0.005 5.137 <0.001 ****
Temperature32:DPE −0.008 0.006 −1.251 0.213 ns
Temperature35:DPE −0.024 0.007 −3.476 <0.001 ***
Phylogenetic
(Intercept) −0.709 0.102 −6.970 <0.001 ****
Temperature32 0.237 0.141 1.682 0.095 ns
Temperature35 0.010 0.144 0.072 ≥0.25 ns
DPE 0.026 0.004 5.907 <0.001 ****
Temperature32:DPE −0.010 0.006 −1.579 0.116 ns
Temperature35:DPE −0.028 0.006 −4.344 <0.001 ****
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Unexposed fish
term statistic df p.value sig
Shannon
Temperature 0.322 2.000 ≥0.25 ns
DPE 22.895 1.000 <0.001 ****
Temperature:DPE 0.363 2.000 ≥0.25 ns
Simpson
Temperature 8.551 2.000 0.014 *
DPE 28.161 1.000 <0.001 ****
Temperature:DPE 0.118 2.000 ≥0.25 ns
Richness
Temperature 31.759 2.000 <0.001 ****
DPE 27.201 1.000 <0.001 ****
Temperature:DPE 12.460 2.000 0.002 **
Phylogenetic
Temperature 38.358 2.000 <0.001 ****
DPE 31.813 1.000 <0.001 ****
Temperature:DPE 19.500 2.000 <0.001 ****
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Unexposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score DPE 14 0 1.065 0.294 3.625 0.003 DPE 28
28 Alpha.Score DPE 21 0 0.722 0.281 2.566 0.076 DPE 28
28 Alpha.Score DPE 28 0 1.187 0.274 4.335 <0.001 DPE 28
28 Alpha.Score DPE 42 0 0.548 0.296 1.853 ≥0.25 DPE 28
28 Alpha.Score DPE 21 14 −0.343 0.317 −1.083 ≥0.25 DPE 28
28 Alpha.Score DPE 28 14 0.122 0.310 0.392 ≥0.25 DPE 28
28 Alpha.Score DPE 42 14 −0.517 0.330 −1.567 ≥0.25 DPE 28
28 Alpha.Score DPE 28 21 0.465 0.299 1.558 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 −0.174 0.319 −0.545 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 −0.639 0.312 −2.045 0.243 DPE 28
32 Alpha.Score DPE 14 0 0.006 0.250 0.022 ≥0.25 DPE 32
32 Alpha.Score DPE 21 0 0.112 0.247 0.452 ≥0.25 DPE 32
32 Alpha.Score DPE 28 0 0.420 0.248 1.695 ≥0.25 DPE 32
32 Alpha.Score DPE 42 0 0.694 0.236 2.943 0.026 DPE 32
32 Alpha.Score DPE 21 14 0.106 0.286 0.371 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.414 0.287 1.445 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 0.688 0.276 2.490 0.092 DPE 32
32 Alpha.Score DPE 28 21 0.308 0.283 1.087 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.582 0.273 2.131 0.205 DPE 32
32 Alpha.Score DPE 42 28 0.274 0.274 1.001 ≥0.25 DPE 32
35 Alpha.Score DPE 14 0 −0.006 0.210 −0.027 ≥0.25 DPE 35
35 Alpha.Score DPE 21 0 −0.249 0.218 −1.145 ≥0.25 DPE 35
35 Alpha.Score DPE 28 0 0.232 0.204 1.138 ≥0.25 DPE 35
35 Alpha.Score DPE 42 0 0.661 0.207 3.194 0.012 DPE 35
35 Alpha.Score DPE 21 14 −0.243 0.253 −0.962 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 0.238 0.242 0.985 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 0.667 0.244 2.733 0.049 DPE 35
35 Alpha.Score DPE 28 21 0.481 0.248 1.938 ≥0.25 DPE 35
35 Alpha.Score DPE 42 21 0.910 0.251 3.628 0.003 DPE 35
35 Alpha.Score DPE 42 28 0.429 0.239 1.792 ≥0.25 DPE 35
Simpson
28 Alpha.Score DPE 14 0 1.256 0.273 4.611 <0.001 DPE 28
28 Alpha.Score DPE 21 0 0.761 0.247 3.076 0.018 DPE 28
28 Alpha.Score DPE 28 0 1.334 0.254 5.241 <0.001 DPE 28
28 Alpha.Score DPE 42 0 0.514 0.257 2.002 ≥0.25 DPE 28
28 Alpha.Score DPE 21 14 −0.496 0.302 −1.644 ≥0.25 DPE 28
28 Alpha.Score DPE 28 14 0.077 0.308 0.251 ≥0.25 DPE 28
28 Alpha.Score DPE 42 14 −0.742 0.310 −2.398 0.114 DPE 28
28 Alpha.Score DPE 28 21 0.573 0.285 2.008 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 −0.246 0.288 −0.857 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 −0.819 0.294 −2.789 0.042 DPE 28
32 Alpha.Score DPE 14 0 0.065 0.249 0.261 ≥0.25 DPE 32
32 Alpha.Score DPE 21 0 0.142 0.249 0.569 ≥0.25 DPE 32
32 Alpha.Score DPE 28 0 0.592 0.261 2.272 0.152 DPE 32
32 Alpha.Score DPE 42 0 0.724 0.254 2.850 0.035 DPE 32
32 Alpha.Score DPE 21 14 0.077 0.287 0.267 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.527 0.297 1.773 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 0.659 0.292 2.260 0.156 DPE 32
32 Alpha.Score DPE 28 21 0.451 0.297 1.517 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.582 0.291 1.999 ≥0.25 DPE 32
32 Alpha.Score DPE 42 28 0.132 0.302 0.436 ≥0.25 DPE 32
35 Alpha.Score DPE 14 0 0.149 0.219 0.683 ≥0.25 DPE 35
35 Alpha.Score DPE 21 0 −0.167 0.218 −0.766 ≥0.25 DPE 35
35 Alpha.Score DPE 28 0 0.413 0.222 1.858 ≥0.25 DPE 35
35 Alpha.Score DPE 42 0 0.836 0.244 3.431 0.005 DPE 35
35 Alpha.Score DPE 21 14 −0.316 0.257 −1.232 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 0.263 0.260 1.013 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 0.687 0.279 2.465 0.097 DPE 35
35 Alpha.Score DPE 28 21 0.579 0.260 2.233 0.165 DPE 35
35 Alpha.Score DPE 42 21 1.003 0.278 3.604 0.003 DPE 35
35 Alpha.Score DPE 42 28 0.424 0.282 1.506 ≥0.25 DPE 35
Richness
28 Alpha.Score DPE 14 0 0.432 0.236 1.836 ≥0.25 DPE 28
28 Alpha.Score DPE 21 0 0.550 0.218 2.526 0.084 DPE 28
28 Alpha.Score DPE 28 0 0.767 0.218 3.525 0.004 DPE 28
28 Alpha.Score DPE 42 0 0.970 0.227 4.278 <0.001 DPE 28
28 Alpha.Score DPE 21 14 0.118 0.263 0.449 ≥0.25 DPE 28
28 Alpha.Score DPE 28 14 0.335 0.262 1.275 ≥0.25 DPE 28
28 Alpha.Score DPE 42 14 0.537 0.270 1.990 ≥0.25 DPE 28
28 Alpha.Score DPE 28 21 0.217 0.247 0.880 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 0.419 0.255 1.647 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 0.203 0.254 0.796 ≥0.25 DPE 28
32 Alpha.Score DPE 14 0 −0.170 0.177 −0.965 ≥0.25 DPE 32
32 Alpha.Score DPE 21 0 −0.026 0.175 −0.147 ≥0.25 DPE 32
32 Alpha.Score DPE 28 0 0.187 0.180 1.041 ≥0.25 DPE 32
32 Alpha.Score DPE 42 0 0.769 0.176 4.359 <0.001 DPE 32
32 Alpha.Score DPE 21 14 0.145 0.204 0.711 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.358 0.208 1.721 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 0.939 0.205 4.585 <0.001 DPE 32
32 Alpha.Score DPE 28 21 0.213 0.206 1.031 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.794 0.203 3.906 <0.001 DPE 32
32 Alpha.Score DPE 42 28 0.581 0.208 2.801 0.040 DPE 32
35 Alpha.Score DPE 14 0 −0.560 0.214 −2.612 0.067 DPE 35
35 Alpha.Score DPE 21 0 −0.551 0.214 −2.578 0.073 DPE 35
35 Alpha.Score DPE 28 0 0.110 0.196 0.561 ≥0.25 DPE 35
35 Alpha.Score DPE 42 0 0.007 0.206 0.036 ≥0.25 DPE 35
35 Alpha.Score DPE 21 14 0.008 0.259 0.032 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 0.670 0.245 2.733 0.048 DPE 35
35 Alpha.Score DPE 42 14 0.567 0.253 2.240 0.162 DPE 35
35 Alpha.Score DPE 28 21 0.661 0.245 2.702 0.052 DPE 35
35 Alpha.Score DPE 42 21 0.559 0.253 2.210 0.173 DPE 35
35 Alpha.Score DPE 42 28 −0.103 0.238 −0.431 ≥0.25 DPE 35
Phylogenetic
28 Alpha.Score DPE 14 0 0.452 0.229 1.969 ≥0.25 DPE 28
28 Alpha.Score DPE 21 0 0.679 0.211 3.210 0.011 DPE 28
28 Alpha.Score DPE 28 0 0.872 0.212 4.120 <0.001 DPE 28
28 Alpha.Score DPE 42 0 1.012 0.220 4.602 <0.001 DPE 28
28 Alpha.Score DPE 21 14 0.227 0.254 0.893 ≥0.25 DPE 28
28 Alpha.Score DPE 28 14 0.420 0.254 1.651 ≥0.25 DPE 28
28 Alpha.Score DPE 42 14 0.560 0.261 2.142 0.200 DPE 28
28 Alpha.Score DPE 28 21 0.193 0.238 0.809 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 0.333 0.246 1.353 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 0.140 0.246 0.569 ≥0.25 DPE 28
32 Alpha.Score DPE 14 0 −0.030 0.164 −0.181 ≥0.25 DPE 32
32 Alpha.Score DPE 21 0 0.154 0.163 0.945 ≥0.25 DPE 32
32 Alpha.Score DPE 28 0 0.260 0.168 1.550 ≥0.25 DPE 32
32 Alpha.Score DPE 42 0 0.753 0.164 4.594 <0.001 DPE 32
32 Alpha.Score DPE 21 14 0.183 0.188 0.974 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.290 0.193 1.503 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 0.783 0.190 4.130 <0.001 DPE 32
32 Alpha.Score DPE 28 21 0.107 0.192 0.556 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.599 0.188 3.185 0.013 DPE 32
32 Alpha.Score DPE 42 28 0.493 0.193 2.555 0.078 DPE 32
35 Alpha.Score DPE 14 0 −0.393 0.204 −1.929 ≥0.25 DPE 35
35 Alpha.Score DPE 21 0 −0.453 0.206 −2.198 0.177 DPE 35
35 Alpha.Score DPE 28 0 0.090 0.192 0.468 ≥0.25 DPE 35
35 Alpha.Score DPE 42 0 −0.111 0.204 −0.542 ≥0.25 DPE 35
35 Alpha.Score DPE 21 14 −0.059 0.246 −0.241 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 0.483 0.235 2.057 0.235 DPE 35
35 Alpha.Score DPE 42 14 0.282 0.245 1.153 ≥0.25 DPE 35
35 Alpha.Score DPE 28 21 0.542 0.237 2.292 0.145 DPE 35
35 Alpha.Score DPE 42 21 0.342 0.247 1.386 ≥0.25 DPE 35
35 Alpha.Score DPE 42 28 −0.200 0.235 −0.853 ≥0.25 DPE 35
Addl. Metrics

2D)

Tables
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ADONIS2
ADONIS2
adonis2(Beta Distance ~ Temperature*DPE); Unexposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 2.294 0.133 16.443 0.001 ***
DPE 1.000 2.531 0.147 36.284 0.001 ***
Temperature:DPE 2.000 0.896 0.052 6.425 0.001 ***
Residual 165.000 11.509 0.668 NA NA NA
Total 170.000 17.229 1.000 NA NA NA
canberra
Temperature 2.000 3.862 0.099 10.667 0.001 ***
DPE 1.000 3.655 0.093 20.192 0.001 ***
Temperature:DPE 2.000 1.820 0.046 5.026 0.001 ***
Residual 165.000 29.869 0.762 NA NA NA
Total 170.000 39.206 1.000 NA NA NA
gunifrac
Temperature 2.000 1.422 0.102 11.984 0.001 ***
DPE 1.000 2.123 0.153 35.785 0.001 ***
Temperature:DPE 2.000 0.574 0.041 4.840 0.001 ***
Residual 165.000 9.788 0.704 NA NA NA
Total 170.000 13.907 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Unexposed fish
term df sumsq meansq statistic p.value sig
bray
Temp.DPE 14.000 0.290 0.021 2.348 0.006 **
Residual 156.000 1.376 0.009 NA NA NA
canberra
Temp.DPE 14.000 0.250 0.018 3.344 <0.001 ***
Residual 156.000 0.831 0.005 NA NA NA
gunifrac
Temp.DPE 14.000 0.249 0.018 2.986 <0.001 ***
Residual 156.000 0.929 0.006 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*DPE); Unexposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temp.DPE 28°C__14DPE 28°C__0DPE 0.024 −0.112 0.159 ≥0.25 ns
Distance Temp.DPE 28°C__21DPE 28°C__0DPE 0.052 −0.074 0.177 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__0DPE 0.007 −0.118 0.133 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__0DPE −0.017 −0.147 0.113 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__0DPE 0.097 −0.005 0.199 0.085 ns
Distance Temp.DPE 32°C__14DPE 28°C__0DPE 0.041 −0.085 0.166 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__0DPE −0.018 −0.143 0.108 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__0DPE 0.028 −0.102 0.158 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__0DPE 0.017 −0.108 0.142 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__0DPE 0.044 −0.058 0.146 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__0DPE 0.004 −0.126 0.134 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__0DPE −0.004 −0.134 0.126 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__0DPE 0.033 −0.097 0.162 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__0DPE 0.146 0.011 0.282 0.021 *
Distance Temp.DPE 28°C__21DPE 28°C__14DPE 0.028 −0.126 0.181 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE −0.016 −0.170 0.137 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__14DPE −0.040 −0.198 0.117 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__14DPE 0.073 −0.062 0.209 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE 0.017 −0.136 0.171 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.041 −0.195 0.112 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE 0.005 −0.153 0.162 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.006 −0.160 0.147 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__14DPE 0.020 −0.115 0.156 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.019 −0.176 0.138 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.028 −0.185 0.129 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE 0.009 −0.148 0.166 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__14DPE 0.123 −0.039 0.284 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE −0.044 −0.189 0.101 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE −0.068 −0.217 0.081 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__21DPE 0.045 −0.080 0.171 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE −0.011 −0.156 0.134 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE −0.069 −0.214 0.076 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE −0.023 −0.172 0.125 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE −0.034 −0.179 0.110 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__21DPE −0.008 −0.133 0.118 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE −0.047 −0.196 0.102 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE −0.056 −0.205 0.093 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.019 −0.168 0.130 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE 0.095 −0.059 0.248 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE −0.024 −0.173 0.125 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__28DPE 0.089 −0.036 0.215 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__28DPE 0.033 −0.111 0.178 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE −0.025 −0.170 0.120 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE 0.021 −0.128 0.169 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE 0.010 −0.135 0.154 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__28DPE 0.036 −0.089 0.162 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE −0.003 −0.152 0.146 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE −0.012 −0.161 0.137 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE 0.025 −0.124 0.174 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE 0.139 −0.015 0.292 0.125 ns
Distance Temp.DPE 32°C__0DPE 28°C__42DPE 0.114 −0.016 0.243 0.161 ns
Distance Temp.DPE 32°C__14DPE 28°C__42DPE 0.057 −0.091 0.206 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE −0.001 −0.150 0.148 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE 0.045 −0.108 0.197 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE 0.034 −0.115 0.183 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__42DPE 0.061 −0.069 0.191 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE 0.021 −0.131 0.174 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE 0.012 −0.140 0.165 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE 0.049 −0.103 0.202 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__42DPE 0.163 0.006 0.320 0.034 *
Distance Temp.DPE 32°C__14DPE 32°C__0DPE −0.056 −0.182 0.069 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__0DPE −0.114 −0.240 0.011 0.115 ns
Distance Temp.DPE 32°C__28DPE 32°C__0DPE −0.069 −0.199 0.061 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__0DPE −0.080 −0.205 0.046 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__0DPE −0.053 −0.155 0.049 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__0DPE −0.092 −0.222 0.037 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__0DPE −0.101 −0.231 0.029 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__0DPE −0.064 −0.194 0.066 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__0DPE 0.049 −0.086 0.185 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.058 −0.203 0.086 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.013 −0.161 0.136 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.024 −0.168 0.121 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__14DPE 0.003 −0.122 0.129 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.036 −0.185 0.112 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.045 −0.194 0.104 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.008 −0.157 0.141 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__14DPE 0.105 −0.048 0.259 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE 0.046 −0.103 0.194 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE 0.035 −0.110 0.179 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__21DPE 0.061 −0.064 0.187 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE 0.022 −0.127 0.171 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE 0.013 −0.136 0.162 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE 0.050 −0.099 0.199 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE 0.164 0.010 0.317 0.025 *
Distance Temp.DPE 32°C__42DPE 32°C__28DPE −0.011 −0.160 0.138 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__28DPE 0.016 −0.114 0.146 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE −0.024 −0.176 0.129 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE −0.033 −0.185 0.120 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE 0.004 −0.148 0.157 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE 0.118 −0.039 0.275 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__42DPE 0.027 −0.099 0.152 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE −0.013 −0.161 0.136 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE −0.021 −0.170 0.127 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE 0.015 −0.133 0.164 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE 0.129 −0.024 0.283 0.209 ns
Distance Temp.DPE 35°C__14DPE 35°C__0DPE −0.039 −0.169 0.090 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__0DPE −0.048 −0.178 0.082 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__0DPE −0.011 −0.141 0.119 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__0DPE 0.102 −0.033 0.238 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE −0.009 −0.161 0.144 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE 0.028 −0.124 0.181 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE 0.142 −0.016 0.299 0.127 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE 0.037 −0.116 0.190 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE 0.151 −0.007 0.308 0.076 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.114 −0.044 0.271 ≥0.25 ns
canberra
Distance Temp.DPE 28°C__14DPE 28°C__0DPE −0.033 −0.138 0.072 ≥0.25 ns
Distance Temp.DPE 28°C__21DPE 28°C__0DPE −0.005 −0.103 0.092 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__0DPE −0.074 −0.171 0.024 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__0DPE −0.069 −0.170 0.032 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__0DPE 0.035 −0.045 0.115 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__0DPE −0.021 −0.119 0.076 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__0DPE −0.070 −0.167 0.028 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__0DPE −0.056 −0.157 0.045 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__0DPE −0.068 −0.165 0.030 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__0DPE 0.008 −0.071 0.088 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__0DPE −0.041 −0.142 0.060 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__0DPE −0.047 −0.148 0.054 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__0DPE −0.052 −0.153 0.049 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__0DPE 0.045 −0.061 0.150 ≥0.25 ns
Distance Temp.DPE 28°C__21DPE 28°C__14DPE 0.028 −0.091 0.147 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE −0.040 −0.160 0.079 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__14DPE −0.035 −0.158 0.087 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__14DPE 0.068 −0.037 0.174 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE 0.012 −0.107 0.131 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.037 −0.156 0.083 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE −0.023 −0.145 0.099 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.034 −0.154 0.085 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__14DPE 0.042 −0.064 0.147 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.008 −0.130 0.115 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.013 −0.136 0.109 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE −0.019 −0.141 0.103 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__14DPE 0.078 −0.048 0.204 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE −0.068 −0.181 0.044 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE −0.063 −0.179 0.052 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__21DPE 0.040 −0.057 0.138 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE −0.016 −0.128 0.097 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE −0.065 −0.177 0.048 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE −0.051 −0.167 0.065 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE −0.062 −0.175 0.050 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__21DPE 0.014 −0.084 0.111 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE −0.036 −0.151 0.080 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE −0.041 −0.157 0.074 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.047 −0.162 0.069 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE 0.050 −0.070 0.169 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE 0.005 −0.110 0.121 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__28DPE 0.109 0.011 0.206 0.014 *
Distance Temp.DPE 32°C__14DPE 28°C__28DPE 0.052 −0.060 0.165 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE 0.004 −0.109 0.116 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE 0.017 −0.098 0.133 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE 0.006 −0.106 0.119 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__28DPE 0.082 −0.015 0.180 0.206 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE 0.033 −0.083 0.148 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE 0.027 −0.088 0.143 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE 0.021 −0.094 0.137 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE 0.118 −0.001 0.237 0.055 ns
Distance Temp.DPE 32°C__0DPE 28°C__42DPE 0.104 0.003 0.205 0.038 *
Distance Temp.DPE 32°C__14DPE 28°C__42DPE 0.047 −0.068 0.163 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE −0.001 −0.117 0.114 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE 0.012 −0.106 0.131 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE 0.001 −0.115 0.117 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__42DPE 0.077 −0.024 0.178 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE 0.028 −0.091 0.146 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE 0.022 −0.097 0.141 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE 0.016 −0.102 0.135 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__42DPE 0.113 −0.009 0.235 0.104 ns
Distance Temp.DPE 32°C__14DPE 32°C__0DPE −0.056 −0.154 0.041 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__0DPE −0.105 −0.202 −0.007 0.022 *
Distance Temp.DPE 32°C__28DPE 32°C__0DPE −0.091 −0.192 0.010 0.125 ns
Distance Temp.DPE 32°C__42DPE 32°C__0DPE −0.103 −0.200 −0.005 0.029 *
Distance Temp.DPE 35°C__0DPE 32°C__0DPE −0.027 −0.106 0.053 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__0DPE −0.076 −0.177 0.025 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__0DPE −0.082 −0.183 0.019 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__0DPE −0.087 −0.188 0.014 0.175 ns
Distance Temp.DPE 35°C__42DPE 32°C__0DPE 0.009 −0.096 0.115 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.049 −0.161 0.064 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.035 −0.151 0.081 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.046 −0.159 0.066 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__14DPE 0.030 −0.068 0.127 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.020 −0.135 0.096 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.025 −0.141 0.090 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.031 −0.147 0.085 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__14DPE 0.066 −0.054 0.185 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE 0.014 −0.102 0.129 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE 0.002 −0.110 0.115 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__21DPE 0.078 −0.019 0.176 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE 0.029 −0.087 0.145 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE 0.023 −0.092 0.139 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE 0.018 −0.098 0.133 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE 0.114 −0.005 0.234 0.076 ns
Distance Temp.DPE 32°C__42DPE 32°C__28DPE −0.011 −0.127 0.104 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__28DPE 0.065 −0.036 0.166 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE 0.015 −0.103 0.134 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE 0.010 −0.109 0.128 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE 0.004 −0.115 0.123 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE 0.101 −0.022 0.223 0.237 ns
Distance Temp.DPE 35°C__0DPE 32°C__42DPE 0.076 −0.021 0.173 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE 0.027 −0.089 0.142 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE 0.021 −0.095 0.137 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE 0.015 −0.100 0.131 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE 0.112 −0.007 0.231 0.091 ns
Distance Temp.DPE 35°C__14DPE 35°C__0DPE −0.049 −0.150 0.052 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__0DPE −0.055 −0.156 0.046 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__0DPE −0.061 −0.162 0.040 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__0DPE 0.036 −0.069 0.141 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE −0.006 −0.124 0.113 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE −0.011 −0.130 0.107 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE 0.085 −0.037 0.208 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE −0.006 −0.124 0.113 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE 0.091 −0.031 0.213 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.097 −0.026 0.219 ≥0.25 ns
gunifrac
Distance Temp.DPE 28°C__14DPE 28°C__0DPE −0.005 −0.116 0.107 ≥0.25 ns
Distance Temp.DPE 28°C__21DPE 28°C__0DPE 0.038 −0.065 0.141 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__0DPE −0.010 −0.113 0.093 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__0DPE −0.033 −0.140 0.073 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__0DPE 0.067 −0.017 0.151 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__0DPE 0.018 −0.085 0.121 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__0DPE −0.023 −0.126 0.080 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__0DPE −0.014 −0.121 0.093 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__0DPE −0.026 −0.129 0.077 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__0DPE 0.029 −0.055 0.113 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__0DPE −0.015 −0.122 0.092 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__0DPE −0.020 −0.127 0.087 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__0DPE −0.065 −0.171 0.042 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__0DPE 0.091 −0.020 0.202 0.245 ns
Distance Temp.DPE 28°C__21DPE 28°C__14DPE 0.042 −0.084 0.168 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE −0.005 −0.131 0.121 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__14DPE −0.029 −0.158 0.100 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__14DPE 0.072 −0.040 0.183 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE 0.022 −0.104 0.148 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.019 −0.145 0.107 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE −0.009 −0.138 0.120 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.021 −0.147 0.105 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__14DPE 0.033 −0.078 0.145 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.010 −0.139 0.119 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.015 −0.145 0.114 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE −0.060 −0.189 0.069 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__14DPE 0.096 −0.037 0.229 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE −0.047 −0.166 0.072 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE −0.071 −0.193 0.051 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__21DPE 0.029 −0.074 0.132 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE −0.020 −0.139 0.099 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE −0.061 −0.180 0.058 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE −0.051 −0.174 0.071 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE −0.063 −0.182 0.056 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__21DPE −0.009 −0.112 0.094 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE −0.052 −0.175 0.070 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE −0.058 −0.180 0.065 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.102 −0.224 0.020 0.216 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE 0.054 −0.073 0.180 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE −0.024 −0.146 0.098 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__28DPE 0.077 −0.026 0.180 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__28DPE 0.027 −0.092 0.146 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE −0.014 −0.133 0.105 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE −0.004 −0.126 0.118 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE −0.016 −0.135 0.103 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__28DPE 0.038 −0.065 0.142 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE −0.005 −0.127 0.117 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE −0.010 −0.132 0.112 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE −0.055 −0.177 0.067 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE 0.101 −0.025 0.227 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__42DPE 0.100 −0.006 0.207 0.089 ns
Distance Temp.DPE 32°C__14DPE 28°C__42DPE 0.051 −0.071 0.173 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE 0.010 −0.112 0.132 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE 0.020 −0.106 0.145 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE 0.008 −0.114 0.130 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__42DPE 0.062 −0.045 0.169 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE 0.019 −0.107 0.144 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE 0.014 −0.112 0.139 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE −0.031 −0.157 0.094 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__42DPE 0.125 −0.005 0.254 0.071 ns
Distance Temp.DPE 32°C__14DPE 32°C__0DPE −0.049 −0.152 0.054 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__0DPE −0.090 −0.193 0.013 0.155 ns
Distance Temp.DPE 32°C__28DPE 32°C__0DPE −0.081 −0.188 0.026 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__0DPE −0.093 −0.196 0.010 0.130 ns
Distance Temp.DPE 35°C__0DPE 32°C__0DPE −0.038 −0.122 0.046 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__0DPE −0.082 −0.189 0.025 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__0DPE −0.087 −0.194 0.020 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__0DPE −0.132 −0.238 −0.025 0.003 **
Distance Temp.DPE 35°C__42DPE 32°C__0DPE 0.024 −0.087 0.135 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.041 −0.160 0.078 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.031 −0.154 0.091 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.043 −0.162 0.076 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__14DPE 0.011 −0.092 0.114 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.032 −0.155 0.090 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.037 −0.160 0.085 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.082 −0.204 0.040 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__14DPE 0.074 −0.053 0.200 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE 0.010 −0.113 0.132 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE −0.002 −0.121 0.117 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__21DPE 0.052 −0.051 0.155 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE 0.009 −0.114 0.131 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE 0.004 −0.119 0.126 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE −0.041 −0.163 0.081 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE 0.115 −0.011 0.241 0.119 ns
Distance Temp.DPE 32°C__42DPE 32°C__28DPE −0.012 −0.134 0.110 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__28DPE 0.043 −0.064 0.149 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE −0.001 −0.126 0.124 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE −0.006 −0.132 0.119 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE −0.051 −0.176 0.075 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE 0.105 −0.024 0.234 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__42DPE 0.054 −0.049 0.157 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE 0.011 −0.111 0.133 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE 0.006 −0.117 0.128 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE −0.039 −0.161 0.083 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE 0.117 −0.009 0.243 0.102 ns
Distance Temp.DPE 35°C__14DPE 35°C__0DPE −0.044 −0.150 0.063 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__0DPE −0.049 −0.155 0.058 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__0DPE −0.093 −0.200 0.013 0.160 ns
Distance Temp.DPE 35°C__42DPE 35°C__0DPE 0.062 −0.049 0.174 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE −0.005 −0.130 0.120 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE −0.050 −0.175 0.076 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE 0.106 −0.023 0.235 0.244 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE −0.045 −0.170 0.081 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE 0.111 −0.018 0.240 0.180 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.156 0.027 0.285 0.005 **
Addl. Metrics

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S2A

S2B

S2C

S2D

Fig. 3) Infection burden is highest in fish reared at lower water temperatures

Fig. 3 Infection outcomes in zebrafish exposed to Pseudocapillaria tomentosa. (A) Histological sections stained with ### stain in zebrafish exposed to P. tomentosa examined at ### days post exposure. Nematodes in intestinal lumen. Arrow = ###. (B) Infection outcome analysis of fish exposed to P. tomentosa (n = 89) by temperature. Fish reared at 28°C and 32°C water temperatures had significantly different infection burden to fish reared at 35°C water temperature. Only one fish reared at 35°C was identified as being positively infected by wet mount. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level.

3A)

3B)

Tables
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GLM
GLM Results
glm.nb(Total.Worm.Count ~ Temperature); Exposed fish
term estimate std.error statistic p.value p.adj.sig
(Intercept) 1.552 0.316 4.914 <0.001 ****
Temperature32 −0.368 0.431 −0.853 ≥0.25 ns
Temperature35 −5.078 1.080 −4.701 <0.001 ****
ANOVA
ANOVA of GLM
ANOVA(GLM.NB(Total.Worm.Count ~ Temperature), type = 2); Exposed fish
term statistic df p.value
Temperature 55.264 2.000 <0.001
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Total.Worm.Count ~ Temperature); Exposed fish
term .y. group1 group2 estimate std.error df statistic adj.p.value
Temperature Total.Worm.Count 28 32 0.368 0.431 Inf 0.853 ≥0.25
Temperature Total.Worm.Count 28 35 5.078 1.080 Inf 4.701 <0.001
Temperature Total.Worm.Count 32 35 4.710 1.074 Inf 4.386 <0.001
(hide)
Pathology Results
Summary of Infection Outcomes
(Pathology Results)
Pathology.Results Count
28
negative 8
positive 17
32
negative 15
positive 15
35
negative 33
positive 1
Total Worm Counts
Summary of Infection Outcomes
(Total Worm Count)
Group Variable Mean SD Min Max n_Missing
28 Total.Worm.Count 5 5 0 14 0
32 Total.Worm.Count 3 5 0 15 0
35 Total.Worm.Count 0 0 0 1 0
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S3B.1

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GLM
GLM Results
glm.nb(Total.Worm.Count ~ Temperature*DPE); Exposed fish
term estimate std.error statistic p.value p.adj.sig
(Intercept) 0.535 0.795 0.673 ≥0.25 ns
Temperature32 0.233 1.081 0.216 ≥0.25 ns
Temperature35 −4.474 2.964 −1.509 0.131 ns
DPE 0.035 0.027 1.304 0.192 ns
Temperature32:DPE −0.019 0.038 −0.501 ≥0.25 ns
Temperature35:DPE −0.020 0.099 −0.200 ≥0.25 ns
ANOVA
ANOVA of GLM
ANOVA(GLM.NB(Total.Worm.Count ~ Temperature*DPE), type = 2); Exposed fish
term statistic df p.value
Temperature 56.284 2.000 <0.001
DPE 1.790 1.000 0.181
Temperature:DPE 0.265 2.000 ≥0.25
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Total.Worm.Count ~ Temperature*DPE); Exposed fish
DPE .y. term group1 group2 estimate std.error df statistic adj.p.value
14 Total.Worm.Count Temperature 28 32 −0.229 0.279 Inf −0.820 ≥0.25
14 Total.Worm.Count Temperature 28 35 38.009 22369621.333 Inf 0.000 ≥0.25
14 Total.Worm.Count Temperature 32 35 38.238 22369621.333 Inf 0.000 ≥0.25
21 Total.Worm.Count Temperature 28 32 0.560 0.392 Inf 1.427 ≥0.25
21 Total.Worm.Count Temperature 28 35 38.222 23726566.406 Inf 0.000 ≥0.25
21 Total.Worm.Count Temperature 32 35 37.663 23726566.406 Inf 0.000 ≥0.25
28 Total.Worm.Count Temperature 28 32 0.605 0.582 Inf 1.040 ≥0.25
28 Total.Worm.Count Temperature 28 35 3.837 1.121 Inf 3.422 0.002
28 Total.Worm.Count Temperature 32 35 3.232 1.119 Inf 2.888 0.011
42 Total.Worm.Count Temperature 28 32 0.377 0.620 Inf 0.609 ≥0.25
42 Total.Worm.Count Temperature 28 35 21.230 3142.206 Inf 0.007 ≥0.25
42 Total.Worm.Count Temperature 32 35 20.853 3142.206 Inf 0.007 ≥0.25

S3B.2

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GLM
GLM Results
glm.nb(Total.Worm.Count ~ Temperature*DPE); Exposed fish
term estimate std.error statistic p.value p.adj.sig
(Intercept) 0.535 0.795 0.673 ≥0.25 ns
Temperature32 0.233 1.081 0.216 ≥0.25 ns
Temperature35 −4.474 2.964 −1.509 0.131 ns
DPE 0.035 0.027 1.304 0.192 ns
Temperature32:DPE −0.019 0.038 −0.501 ≥0.25 ns
Temperature35:DPE −0.020 0.099 −0.200 ≥0.25 ns
ANOVA
ANOVA of GLM
ANOVA(GLM.NB(Total.Worm.Count ~ Temperature*DPE), type = 2); Exposed fish
term statistic df p.value
Temperature 56.284 2.000 <0.001
DPE 1.790 1.000 0.181
Temperature:DPE 0.265 2.000 ≥0.25

S3C

(hide)
Counts
Detection Method Comparison by DPE
Number of positive and negative detections for Wet and Histo methods
Days Post Exposure Histo Positive Wet Positive Histo Negative Wet Negative
28
14 9 6 0 4
21 6 6 2 4
28 6 8 0 0
42 8 7 1 2
32
14 6 4 3 6
21 5 5 0 5
28 7 7 1 2
42 7 7 2 2
35
14 1 1 9 9
21 6 0 3 10
28 1 1 2 7
42 1 0 9 10
NA
NA 0 0 0 0
Stats
Statistical Comparison of Detection Methods
McNemar's test comparing Wet and Histo methods by Temperature and DPE
Days Post Exposure Histo Only Wet Only Both Positive Both Negative P-value1 Chi-squared
28
14 3 0 6 0 0.2482 1.333333
21 1 0 5 2 1.0000 0.000000
28 0 0 6 0 NaN NaN
42 1 0 6 1 1.0000 0.000000
32
14 4 1 2 2 0.3711 0.800000
21 2 0 3 0 0.4795 0.500000
28 1 0 6 1 1.0000 0.000000
42 2 2 5 0 1.0000 0.000000
35
14 1 1 0 8 1.0000 0.000000
21 6 0 0 3 0.0412 4.166667
28 0 0 1 2 NaN NaN
42 1 0 0 9 1.0000 0.000000
NA
NA 0 0 0 0 NaN NaN
1 McNemar's test compares the discordant pairs (Wet Only vs Histo Only)

Fig. 4) Gut microbiome response to parasite exposure varies across water temperature

Fig. 4 Effects of Pseudocapillaria tomentosa exposure on zebrafish gut microbiomes reared at different water temperatures. (A) Simpson’s Index of diversity shows that gut microbiome diversity significantly differs between fish reared at 28°C water temperature to fish reared at 32°C and 35°C water temperatures. (B) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main effect of temperature. The analysis shows that gut microbiome composition significantly differs between parasite exposed fish reared at different water temperatures. (C) Simpson’s Index of diversity shows microbial gut diversity decreases with time from 0 days post exposure (dpe) to 42 dpe in parasite exposed fish reared at 28°C water temperature. (D) Capscale ordination of gut microbiome composition based on the Canberra dissimilarity constrained on the main effects of water temperature and time (days post exposure, dpe), and their interaction. The analysis shows that shows that gut microbiome composition differs between parasite exposed fish across time depending on water temperature. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate direction of greatest change in the indicated covariates.

4A)

Tables
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GLM
GLM Results
glm(Alpha.Score ~ Temperature); Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −0.936 0.233 −4.021 <0.001 ***
Temperature32 0.909 0.301 3.017 0.003 **
Temperature35 0.516 0.296 1.741 0.085 ns
Simpson
(Intercept) −1.002 0.243 −4.130 <0.001 ****
Temperature32 1.057 0.312 3.384 0.001 **
Temperature35 0.772 0.306 2.525 0.013 *
Richness
(Intercept) −0.681 0.190 −3.583 <0.001 ***
Temperature32 0.703 0.251 2.800 0.006 **
Temperature35 0.242 0.247 0.978 ≥0.25 ns
Phylogenetic
(Intercept) −0.681 0.180 −3.789 <0.001 ***
Temperature32 0.648 0.237 2.730 0.008 **
Temperature35 0.161 0.235 0.688 ≥0.25 ns
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature), type = 2); Exposed fish
term statistic df p.value sig
Shannon
Temperature 9.440 2.000 0.009 **
Simpson
Temperature 12.497 2.000 0.002 **
Richness
Temperature 8.575 2.000 0.014 *
Phylogenetic
Temperature 8.706 2.000 0.013 *
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature); Exposed fish
term .y. group1 group2 estimate std.error statistic adj.p.value Variable
Shannon
Temperature Alpha.Score 32 28 0.909 0.301 3.017 0.007 Temperature
Temperature Alpha.Score 35 28 0.516 0.296 1.741 0.190 Temperature
Temperature Alpha.Score 35 32 −0.393 0.265 −1.482 ≥0.25 Temperature
Simpson
Temperature Alpha.Score 32 28 1.057 0.312 3.384 0.002 Temperature
Temperature Alpha.Score 35 28 0.772 0.306 2.525 0.031 Temperature
Temperature Alpha.Score 35 32 −0.285 0.270 −1.054 ≥0.25 Temperature
Richness
Temperature Alpha.Score 32 28 0.703 0.251 2.800 0.014 Temperature
Temperature Alpha.Score 35 28 0.242 0.247 0.978 ≥0.25 Temperature
Temperature Alpha.Score 35 32 −0.461 0.228 −2.028 0.105 Temperature
Phylogenetic
Temperature Alpha.Score 32 28 0.648 0.237 2.730 0.017 Temperature
Temperature Alpha.Score 35 28 0.161 0.235 0.688 ≥0.25 Temperature
Temperature Alpha.Score 35 32 −0.487 0.216 −2.251 0.063 Temperature
Addl. Metrics

4B)

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ADONIS2
ADONIS2
adonis2(Beta Distance ~ Temperature); Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 5.218 0.251 14.447 0.001 ***
Residual 86.000 15.530 0.749 NA NA NA
Total 88.000 20.748 1.000 NA NA NA
canberra
Temperature 2.000 3.573 0.117 5.701 0.001 ***
Residual 86.000 26.947 0.883 NA NA NA
Total 88.000 30.520 1.000 NA NA NA
gunifrac
Temperature 2.000 3.363 0.212 11.590 0.001 ***
Residual 86.000 12.478 0.788 NA NA NA
Total 88.000 15.841 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Exposed fish
term df sumsq meansq statistic p.value sig
bray
Temperature 2.000 0.364 0.182 8.708 <0.001 ***
Residual 86.000 1.796 0.021 NA NA NA
canberra
Temperature 2.000 0.314 0.157 17.419 <0.001 ****
Residual 86.000 0.774 0.009 NA NA NA
gunifrac
Temperature 2.000 0.310 0.155 13.567 <0.001 ****
Residual 86.000 0.983 0.011 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature); Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temperature 32 28 −0.043 −0.136 0.051 ≥0.25 ns
Distance Temperature 35 28 −0.150 −0.241 −0.059 <0.001 ***
Distance Temperature 35 32 −0.108 −0.194 −0.021 0.011 *
canberra
Distance Temperature 32 28 −0.110 −0.171 −0.049 <0.001 ***
Distance Temperature 35 28 −0.144 −0.204 −0.085 <0.001 ****
Distance Temperature 35 32 −0.034 −0.091 0.022 ≥0.25 ns
gunifrac
Distance Temperature 32 28 −0.056 −0.125 0.013 0.133 ns
Distance Temperature 35 28 −0.143 −0.211 −0.076 <0.001 ****
Distance Temperature 35 32 −0.087 −0.151 −0.023 0.005 **
Addl. Metrics

4C)

Tables
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GLM
GLM Results
glm(Alpha.Score ~ Temperature*DPE); Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −0.134 0.594 −0.226 ≥0.25 ns
Temperature32 0.228 0.774 0.295 ≥0.25 ns
Temperature35 −0.428 0.777 −0.551 ≥0.25 ns
DPE −0.031 0.022 −1.425 0.158 ns
Temperature32:DPE 0.026 0.028 0.917 ≥0.25 ns
Temperature35:DPE 0.036 0.028 1.300 0.197 ns
Simpson
(Intercept) −0.282 0.617 −0.456 ≥0.25 ns
Temperature32 0.336 0.801 0.420 ≥0.25 ns
Temperature35 −0.172 0.799 −0.215 ≥0.25 ns
DPE −0.028 0.022 −1.234 0.221 ns
Temperature32:DPE 0.028 0.029 0.946 ≥0.25 ns
Temperature35:DPE 0.036 0.029 1.261 0.211 ns
Richness
(Intercept) 0.316 0.480 0.657 ≥0.25 ns
Temperature32 −0.034 0.636 −0.054 ≥0.25 ns
Temperature35 −0.596 0.638 −0.934 ≥0.25 ns
DPE −0.038 0.017 −2.191 0.031 *
Temperature32:DPE 0.028 0.023 1.199 0.234 ns
Temperature35:DPE 0.032 0.023 1.405 0.164 ns
Phylogenetic
(Intercept) 0.305 0.451 0.676 ≥0.25 ns
Temperature32 −0.101 0.597 −0.169 ≥0.25 ns
Temperature35 −0.766 0.601 −1.274 0.206 ns
DPE −0.038 0.016 −2.310 0.023 *
Temperature32:DPE 0.028 0.022 1.298 0.198 ns
Temperature35:DPE 0.036 0.022 1.651 0.103 ns
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Exposed fish
term statistic df p.value sig
Shannon
Temperature 8.789 2.000 0.012 *
DPE 0.523 1.000 ≥0.25 ns
Temperature:DPE 1.759 2.000 ≥0.25 ns
Simpson
Temperature 11.871 2.000 0.003 **
DPE 0.120 1.000 ≥0.25 ns
Temperature:DPE 1.684 2.000 ≥0.25 ns
Richness
Temperature 7.911 2.000 0.019 *
DPE 3.435 1.000 0.064 ns
Temperature:DPE 2.255 2.000 ≥0.25 ns
Phylogenetic
Temperature 8.173 2.000 0.017 *
DPE 3.103 1.000 0.078 ns
Temperature:DPE 2.980 2.000 0.225 ns
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Exposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score DPE 21 14 −3.311 1.822 −1.817 0.246 DPE 28
28 Alpha.Score DPE 28 14 −2.027 1.023 −1.982 0.178 DPE 28
28 Alpha.Score DPE 42 14 −1.024 0.705 −1.452 ≥0.25 DPE 28
28 Alpha.Score DPE 28 21 1.284 1.982 0.648 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 2.287 1.838 1.245 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 1.004 1.050 0.956 ≥0.25 DPE 28
32 Alpha.Score DPE 21 14 −0.536 0.537 −0.999 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.056 0.558 0.099 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 −0.271 0.532 −0.509 ≥0.25 DPE 32
32 Alpha.Score DPE 28 21 0.592 0.606 0.977 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.266 0.581 0.457 ≥0.25 DPE 32
32 Alpha.Score DPE 42 28 −0.326 0.601 −0.543 ≥0.25 DPE 32
35 Alpha.Score DPE 21 14 −0.246 0.270 −0.909 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 −0.292 0.271 −1.075 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 0.097 0.258 0.377 ≥0.25 DPE 35
35 Alpha.Score DPE 28 21 −0.046 0.283 −0.162 ≥0.25 DPE 35
35 Alpha.Score DPE 42 21 0.343 0.270 1.273 ≥0.25 DPE 35
35 Alpha.Score DPE 42 28 0.389 0.271 1.438 ≥0.25 DPE 35
Simpson
28 Alpha.Score DPE 21 14 −3.141 1.820 −1.725 ≥0.25 DPE 28
28 Alpha.Score DPE 28 14 −1.994 1.072 −1.861 0.227 DPE 28
28 Alpha.Score DPE 42 14 −0.925 0.721 −1.283 ≥0.25 DPE 28
28 Alpha.Score DPE 28 21 1.147 2.001 0.573 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 2.215 1.837 1.206 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 1.069 1.100 0.971 ≥0.25 DPE 28
32 Alpha.Score DPE 21 14 −0.416 0.550 −0.756 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.275 0.580 0.474 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 −0.138 0.546 −0.252 ≥0.25 DPE 32
32 Alpha.Score DPE 28 21 0.691 0.626 1.103 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.278 0.595 0.467 ≥0.25 DPE 32
32 Alpha.Score DPE 42 28 −0.413 0.623 −0.662 ≥0.25 DPE 32
35 Alpha.Score DPE 21 14 −0.374 0.297 −1.258 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 −0.285 0.296 −0.966 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 0.142 0.284 0.502 ≥0.25 DPE 35
35 Alpha.Score DPE 28 21 0.088 0.308 0.287 ≥0.25 DPE 35
35 Alpha.Score DPE 42 21 0.516 0.297 1.738 ≥0.25 DPE 35
35 Alpha.Score DPE 42 28 0.428 0.295 1.448 ≥0.25 DPE 35
Richness
28 Alpha.Score DPE 21 14 −2.219 1.016 −2.185 0.123 DPE 28
28 Alpha.Score DPE 28 14 −1.585 0.795 −1.993 0.184 DPE 28
28 Alpha.Score DPE 42 14 −1.239 0.650 −1.907 0.218 DPE 28
28 Alpha.Score DPE 28 21 0.634 1.133 0.560 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 0.980 1.036 0.946 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 0.346 0.821 0.421 ≥0.25 DPE 28
32 Alpha.Score DPE 21 14 −0.561 0.412 −1.363 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 −0.470 0.430 −1.091 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 −0.357 0.410 −0.871 ≥0.25 DPE 32
32 Alpha.Score DPE 28 21 0.092 0.462 0.199 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.204 0.444 0.460 ≥0.25 DPE 32
32 Alpha.Score DPE 42 28 0.112 0.461 0.243 ≥0.25 DPE 32
35 Alpha.Score DPE 21 14 −0.338 0.242 −1.396 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 −0.329 0.242 −1.361 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 −0.233 0.233 −1.000 ≥0.25 DPE 35
35 Alpha.Score DPE 28 21 0.009 0.252 0.035 ≥0.25 DPE 35
35 Alpha.Score DPE 42 21 0.105 0.244 0.430 ≥0.25 DPE 35
35 Alpha.Score DPE 42 28 0.096 0.244 0.394 ≥0.25 DPE 35
Phylogenetic
28 Alpha.Score DPE 21 14 −2.030 0.921 −2.205 0.118 DPE 28
28 Alpha.Score DPE 28 14 −1.472 0.742 −1.985 0.188 DPE 28
28 Alpha.Score DPE 42 14 −1.216 0.616 −1.975 0.192 DPE 28
28 Alpha.Score DPE 28 21 0.557 1.029 0.542 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 0.814 0.942 0.864 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 0.256 0.768 0.334 ≥0.25 DPE 28
32 Alpha.Score DPE 21 14 −0.537 0.382 −1.406 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 −0.442 0.399 −1.108 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 −0.329 0.380 −0.866 ≥0.25 DPE 32
32 Alpha.Score DPE 28 21 0.095 0.430 0.222 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.208 0.413 0.504 ≥0.25 DPE 32
32 Alpha.Score DPE 42 28 0.113 0.428 0.263 ≥0.25 DPE 32
35 Alpha.Score DPE 21 14 −0.273 0.236 −1.159 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 −0.214 0.235 −0.911 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 −0.121 0.226 −0.535 ≥0.25 DPE 35
35 Alpha.Score DPE 28 21 0.059 0.246 0.242 ≥0.25 DPE 35
35 Alpha.Score DPE 42 21 0.152 0.237 0.642 ≥0.25 DPE 35
35 Alpha.Score DPE 42 28 0.093 0.236 0.393 ≥0.25 DPE 35
Addl. Metrics

4D)

Tables
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ADONIS2
ADONIS2
adonis2(Beta Distance ~ Temperature*DPE); Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 5.218 0.251 14.843 0.001 ***
DPE 1.000 0.394 0.019 2.241 0.056 ns
Temperature:DPE 2.000 0.548 0.026 1.558 0.113 ns
Residual 83.000 14.589 0.703 NA NA NA
Total 88.000 20.748 1.000 NA NA NA
canberra
Temperature 2.000 3.573 0.117 5.847 0.001 ***
DPE 1.000 0.674 0.022 2.207 0.006 **
Temperature:DPE 2.000 0.913 0.030 1.494 0.017 *
Residual 83.000 25.360 0.831 NA NA NA
Total 88.000 30.520 1.000 NA NA NA
gunifrac
Temperature 2.000 3.363 0.212 11.847 0.001 ***
DPE 1.000 0.261 0.016 1.837 0.101 ns
Temperature:DPE 2.000 0.436 0.028 1.535 0.102 ns
Residual 83.000 11.781 0.744 NA NA NA
Total 88.000 15.841 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Exposed fish
term df sumsq meansq statistic p.value sig
bray
Temp.DPE 11.000 0.624 0.057 3.261 0.001 **
Residual 77.000 1.339 0.017 NA NA NA
canberra
Temp.DPE 11.000 0.316 0.029 2.748 0.005 **
Residual 77.000 0.805 0.010 NA NA NA
gunifrac
Temp.DPE 11.000 0.420 0.038 3.232 0.001 **
Residual 77.000 0.909 0.012 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*DPE); Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temp.DPE 28°C__21DPE 28°C__14DPE −0.213 −0.485 0.059 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE −0.258 −0.512 −0.005 0.042 *
Distance Temp.DPE 28°C__42DPE 28°C__14DPE −0.063 −0.285 0.160 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE −0.059 −0.270 0.152 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.089 −0.319 0.141 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE −0.123 −0.363 0.117 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.142 −0.372 0.088 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.202 −0.418 0.014 0.090 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.170 −0.393 0.052 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE −0.289 −0.511 −0.067 0.002 **
Distance Temp.DPE 35°C__42DPE 28°C__14DPE −0.192 −0.408 0.024 0.130 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE −0.046 −0.344 0.253 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE 0.150 −0.122 0.422 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE 0.153 −0.110 0.416 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE 0.124 −0.154 0.403 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE 0.090 −0.197 0.377 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE 0.071 −0.208 0.350 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE 0.011 −0.256 0.278 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE 0.042 −0.230 0.315 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.077 −0.349 0.196 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE 0.021 −0.246 0.288 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE 0.196 −0.058 0.449 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__28DPE 0.199 −0.044 0.443 0.220 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE 0.170 −0.091 0.430 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE 0.136 −0.133 0.405 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE 0.117 −0.144 0.377 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE 0.057 −0.191 0.305 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE 0.088 −0.165 0.341 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE −0.031 −0.284 0.222 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE 0.066 −0.182 0.314 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__42DPE 0.003 −0.207 0.214 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE −0.026 −0.256 0.204 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE −0.060 −0.300 0.180 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE −0.079 −0.309 0.151 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE −0.139 −0.355 0.077 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE −0.108 −0.330 0.115 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE −0.227 −0.449 −0.004 0.042 *
Distance Temp.DPE 35°C__42DPE 28°C__42DPE −0.129 −0.345 0.087 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.029 −0.248 0.190 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.063 −0.293 0.166 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.082 −0.301 0.137 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.142 −0.347 0.062 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.111 −0.322 0.100 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.230 −0.441 −0.019 0.021 *
Distance Temp.DPE 35°C__42DPE 32°C__14DPE −0.133 −0.337 0.072 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE −0.034 −0.281 0.213 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE −0.053 −0.291 0.185 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE −0.113 −0.337 0.111 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE −0.082 −0.312 0.148 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE −0.201 −0.431 0.029 0.148 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE −0.103 −0.327 0.121 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__28DPE −0.019 −0.266 0.228 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE −0.079 −0.313 0.155 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE −0.048 −0.288 0.192 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE −0.167 −0.407 0.073 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE −0.069 −0.304 0.165 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE −0.060 −0.284 0.164 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE −0.029 −0.259 0.201 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE −0.148 −0.378 0.082 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE −0.050 −0.274 0.174 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE 0.031 −0.185 0.247 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE −0.088 −0.304 0.128 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE 0.010 −0.200 0.219 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE −0.119 −0.341 0.103 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE −0.022 −0.238 0.194 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.097 −0.119 0.313 ≥0.25 ns
canberra
Distance Temp.DPE 28°C__21DPE 28°C__14DPE −0.004 −0.215 0.207 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE 0.040 −0.156 0.237 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__14DPE 0.059 −0.113 0.232 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE −0.043 −0.206 0.121 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.044 −0.223 0.134 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE −0.102 −0.288 0.084 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.084 −0.262 0.095 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.097 −0.265 0.070 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.097 −0.269 0.075 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE −0.142 −0.314 0.031 0.215 ns
Distance Temp.DPE 35°C__42DPE 28°C__14DPE −0.103 −0.271 0.064 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE 0.044 −0.187 0.276 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE 0.064 −0.148 0.275 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE −0.039 −0.243 0.165 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE −0.040 −0.256 0.176 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE −0.097 −0.320 0.125 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE −0.079 −0.295 0.137 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE −0.093 −0.300 0.114 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE −0.093 −0.304 0.118 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.137 −0.348 0.074 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE −0.099 −0.306 0.108 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE 0.019 −0.177 0.216 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__28DPE −0.083 −0.272 0.106 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE −0.085 −0.286 0.117 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE −0.142 −0.351 0.067 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE −0.124 −0.326 0.078 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE −0.137 −0.329 0.055 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE −0.137 −0.334 0.059 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE −0.182 −0.378 0.015 0.097 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE −0.144 −0.336 0.049 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__42DPE −0.102 −0.266 0.061 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE −0.104 −0.282 0.075 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE −0.161 −0.347 0.025 0.157 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE −0.143 −0.321 0.036 0.247 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE −0.156 −0.324 0.011 0.091 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE −0.156 −0.329 0.016 0.112 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE −0.201 −0.373 −0.028 0.010 **
Distance Temp.DPE 35°C__42DPE 28°C__42DPE −0.163 −0.330 0.005 0.065 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.001 −0.171 0.168 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.059 −0.237 0.119 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.041 −0.211 0.129 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.054 −0.212 0.104 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.054 −0.218 0.109 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.099 −0.262 0.065 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__14DPE −0.060 −0.219 0.098 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE −0.057 −0.249 0.134 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE −0.039 −0.224 0.145 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE −0.053 −0.226 0.121 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE −0.053 −0.231 0.126 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE −0.097 −0.276 0.081 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE −0.059 −0.233 0.115 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__28DPE 0.018 −0.174 0.210 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE 0.005 −0.177 0.186 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE 0.005 −0.182 0.191 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE −0.040 −0.226 0.146 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE −0.002 −0.183 0.180 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE −0.013 −0.187 0.160 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE −0.014 −0.192 0.165 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE −0.058 −0.236 0.120 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE −0.020 −0.194 0.154 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE 0.000 −0.168 0.167 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE −0.044 −0.212 0.123 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE −0.006 −0.169 0.156 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE −0.044 −0.217 0.128 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE −0.006 −0.174 0.161 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.038 −0.129 0.206 ≥0.25 ns
gunifrac
Distance Temp.DPE 28°C__21DPE 28°C__14DPE −0.191 −0.415 0.033 0.173 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE −0.061 −0.270 0.147 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__14DPE 0.018 −0.165 0.201 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE −0.024 −0.197 0.150 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.027 −0.216 0.163 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE −0.100 −0.298 0.098 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.069 −0.259 0.120 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.131 −0.309 0.047 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.137 −0.320 0.046 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE −0.198 −0.382 −0.015 0.022 *
Distance Temp.DPE 35°C__42DPE 28°C__14DPE −0.134 −0.312 0.044 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE 0.130 −0.116 0.375 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE 0.209 −0.015 0.433 0.092 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE 0.167 −0.049 0.384 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE 0.164 −0.065 0.394 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE 0.091 −0.146 0.327 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE 0.122 −0.108 0.351 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE 0.060 −0.161 0.280 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE 0.054 −0.170 0.278 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.007 −0.232 0.217 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE 0.057 −0.163 0.277 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE 0.079 −0.130 0.288 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__28DPE 0.038 −0.163 0.238 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE 0.035 −0.180 0.249 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE −0.039 −0.260 0.183 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE −0.008 −0.222 0.206 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE −0.070 −0.274 0.134 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE −0.075 −0.284 0.133 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE −0.137 −0.346 0.072 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE −0.072 −0.277 0.132 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__42DPE −0.041 −0.215 0.132 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE −0.045 −0.234 0.145 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE −0.118 −0.316 0.080 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE −0.087 −0.277 0.102 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE −0.149 −0.327 0.029 0.190 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE −0.155 −0.338 0.029 0.182 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE −0.216 −0.399 −0.033 0.008 **
Distance Temp.DPE 35°C__42DPE 28°C__42DPE −0.151 −0.329 0.026 0.173 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.003 −0.184 0.177 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.076 −0.266 0.113 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.046 −0.226 0.135 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.108 −0.276 0.060 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.113 −0.287 0.061 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.175 −0.348 −0.001 0.047 *
Distance Temp.DPE 35°C__42DPE 32°C__14DPE −0.110 −0.278 0.058 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE −0.073 −0.277 0.130 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE −0.043 −0.238 0.153 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE −0.105 −0.289 0.080 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE −0.110 −0.300 0.079 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE −0.172 −0.361 0.018 0.113 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE −0.107 −0.292 0.078 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__28DPE 0.031 −0.173 0.234 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE −0.031 −0.224 0.162 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE −0.037 −0.234 0.161 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE −0.098 −0.296 0.099 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE −0.034 −0.227 0.159 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE −0.062 −0.247 0.122 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE −0.067 −0.257 0.122 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE −0.129 −0.319 0.060 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE −0.064 −0.249 0.120 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE −0.005 −0.183 0.173 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE −0.067 −0.245 0.111 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE −0.002 −0.175 0.170 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE −0.062 −0.245 0.121 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE 0.003 −0.175 0.181 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.065 −0.113 0.243 ≥0.25 ns
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S4A

S4B

S4C

S4D

Fig. 5) Gut microbiome response has a non-linear relationship with infection burden

Fig. 5 The impacts of presence of infection and infection burden on the gut microbiomes of Pseudocapillaria tomentosa exposed zebrafish. (A) Simpson’s Index for diversity of parasite exposed fish. Gut microbial alpha-diversity does not significantly differ between fish reared at the same water temperature depending on presence of infection. (B) Capscale ordination based on the Canberra dissimilarity of gut microbiome composition of parasite exposed fish constrained on the main effects of temperature and pathology result. The analysis shows that gut microbiome composition significantly differs between positively infected fish reared at different water temperatures. (C) Infection burden (total worm counts ) is positively correlated with lowest or highest alpha diversity scores in positively infected fish. (D) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main effects of water temperature and infection burden. The analysis shows that gut microbiome composition significantly differs between clusters of Low, High and Other fish. Samples points are colored by water temperature, and filled by cluster grouping. Samples with at least one detectable worm and an alpha-diversity score less than 0.5 are categorized as Low (orange fill), samples with at least one detectable worm and an alpha-diversity score greater than 0.5 are categorized as High (purple fill), and samples with no observable infection are categorized as Other (white and transparent fill). Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate statistically significant covariates and direction of greatest change in the indicated covariates.

5A)

Tables
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GLM
GLM Results
glm(Alpha.Score ~ Temperature*Pathology.Results); Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −1.090 0.429 −2.544 0.013 *
Temperature32 0.889 0.508 1.749 0.084 ns
Temperature35 0.693 0.468 1.482 0.142 ns
Pathology.Resultspositive 0.223 0.512 0.436 ≥0.25 ns
Temperature32:Pathology.Resultspositive 0.125 0.641 0.195 ≥0.25 ns
Temperature35:Pathology.Resultspositive −1.122 1.392 −0.806 ≥0.25 ns
Simpson
(Intercept) −1.265 0.458 −2.760 0.007 **
Temperature32 1.105 0.536 2.061 0.042 *
Temperature35 1.055 0.495 2.130 0.036 *
Pathology.Resultspositive 0.375 0.541 0.694 ≥0.25 ns
Temperature32:Pathology.Resultspositive 0.054 0.669 0.081 ≥0.25 ns
Temperature35:Pathology.Resultspositive −1.154 1.337 −0.863 ≥0.25 ns
Richness
(Intercept) −0.421 0.329 −1.279 0.204 ns
Temperature32 0.483 0.405 1.194 0.236 ns
Temperature35 −0.002 0.367 −0.005 ≥0.25 ns
Pathology.Resultspositive −0.390 0.407 −0.957 ≥0.25 ns
Temperature32:Pathology.Resultspositive 0.310 0.526 0.589 ≥0.25 ns
Temperature35:Pathology.Resultspositive −0.221 1.125 −0.197 ≥0.25 ns
Phylogenetic
(Intercept) −0.421 0.311 −1.353 0.180 ns
Temperature32 0.397 0.383 1.037 ≥0.25 ns
Temperature35 −0.085 0.348 −0.244 ≥0.25 ns
Pathology.Resultspositive −0.390 0.385 −1.014 ≥0.25 ns
Temperature32:Pathology.Resultspositive 0.373 0.497 0.750 ≥0.25 ns
Temperature35:Pathology.Resultspositive −0.112 1.058 −0.105 ≥0.25 ns
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Exposed fish
term statistic df p.value sig
Shannon
Temperature 9.895 2.000 0.007 **
Pathology.Results 0.569 1.000 ≥0.25 ns
Temperature:Pathology.Results 0.997 2.000 ≥0.25 ns
Simpson
Temperature 13.663 2.000 0.001 **
Pathology.Results 1.134 1.000 ≥0.25 ns
Temperature:Pathology.Results 0.999 2.000 ≥0.25 ns
Richness
Temperature 8.625 2.000 0.013 *
Pathology.Results 0.840 1.000 ≥0.25 ns
Temperature:Pathology.Results 0.495 2.000 ≥0.25 ns
Phylogenetic
Temperature 8.874 2.000 0.012 *
Pathology.Results 0.625 1.000 ≥0.25 ns
Temperature:Pathology.Results 0.674 2.000 ≥0.25 ns
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Exposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score Pathology.Results positive negative 0.223 0.732 0.305 ≥0.25 Pathology.Results 28
32 Alpha.Score Pathology.Results positive negative 0.348 0.383 0.908 ≥0.25 Pathology.Results 32
35 Alpha.Score Pathology.Results positive negative −0.899 0.659 −1.365 0.172 Pathology.Results 35
Simpson
28 Alpha.Score Pathology.Results positive negative 0.375 0.754 0.497 ≥0.25 Pathology.Results 28
32 Alpha.Score Pathology.Results positive negative 0.429 0.394 1.089 ≥0.25 Pathology.Results 32
35 Alpha.Score Pathology.Results positive negative −0.779 0.692 −1.125 ≥0.25 Pathology.Results 35
Richness
28 Alpha.Score Pathology.Results positive negative −0.390 0.608 −0.641 ≥0.25 Pathology.Results 28
32 Alpha.Score Pathology.Results positive negative −0.080 0.301 −0.267 ≥0.25 Pathology.Results 32
35 Alpha.Score Pathology.Results positive negative −0.611 0.552 −1.107 ≥0.25 Pathology.Results 35
Phylogenetic
28 Alpha.Score Pathology.Results positive negative −0.390 0.577 −0.676 ≥0.25 Pathology.Results 28
32 Alpha.Score Pathology.Results positive negative −0.017 0.280 −0.062 ≥0.25 Pathology.Results 32
35 Alpha.Score Pathology.Results positive negative −0.502 0.524 −0.958 ≥0.25 Pathology.Results 35
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5B)

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ADONIS2
ADONIS2
adonis2(Beta Distance ~ Temperature*Pathology.Results); Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 5.218 0.251 14.569 0.001 ***
Pathology.Results 1.000 0.263 0.013 1.471 0.179 ns
Temperature:Pathology.Results 2.000 0.404 0.019 1.127 ≥0.25 ns
Residual 83.000 14.863 0.716 NA NA NA
Total 88.000 20.748 1.000 NA NA NA
canberra
Temperature 2.000 3.573 0.117 5.751 0.001 ***
Pathology.Results 1.000 0.488 0.016 1.570 0.029 *
Temperature:Pathology.Results 2.000 0.679 0.022 1.093 ≥0.25 ns
Residual 83.000 25.781 0.845 NA NA NA
Total 88.000 30.520 1.000 NA NA NA
gunifrac
Temperature 2.000 3.363 0.212 11.651 0.001 ***
Pathology.Results 1.000 0.234 0.015 1.622 0.141 ns
Temperature:Pathology.Results 2.000 0.264 0.017 0.915 ≥0.25 ns
Residual 83.000 11.980 0.756 NA NA NA
Total 88.000 15.841 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature*Pathology.Results); Exposed fish
term df sumsq meansq statistic p.value sig
bray
Temp.Path 5.000 0.520 0.104 4.652 <0.001 ***
Residual 83.000 1.855 0.022 NA NA NA
canberra
Temp.Path 5.000 0.625 0.125 13.830 <0.001 ****
Residual 83.000 0.750 0.009 NA NA NA
gunifrac
Temp.Path 5.000 0.436 0.087 6.954 <0.001 ****
Residual 83.000 1.040 0.013 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Pathology.Results); Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temp.Path 28°C__pos 28°C__neg 0.022 −0.165 0.209 ≥0.25 ns
Distance Temp.Path 32°C__neg 28°C__neg −0.086 −0.277 0.105 ≥0.25 ns
Distance Temp.Path 32°C__pos 28°C__neg 0.015 −0.176 0.206 ≥0.25 ns
Distance Temp.Path 35°C__neg 28°C__neg −0.126 −0.298 0.046 ≥0.25 ns
Distance Temp.Path 35°C__pos 28°C__neg −0.434 −0.896 0.029 0.079 ns
Distance Temp.Path 32°C__neg 28°C__pos −0.108 −0.263 0.046 ≥0.25 ns
Distance Temp.Path 32°C__pos 28°C__pos −0.007 −0.161 0.148 ≥0.25 ns
Distance Temp.Path 35°C__neg 28°C__pos −0.148 −0.278 −0.017 0.017 *
Distance Temp.Path 35°C__pos 28°C__pos −0.456 −0.905 −0.007 0.044 *
Distance Temp.Path 32°C__pos 32°C__neg 0.101 −0.058 0.261 ≥0.25 ns
Distance Temp.Path 35°C__neg 32°C__neg −0.039 −0.175 0.096 ≥0.25 ns
Distance Temp.Path 35°C__pos 32°C__neg −0.348 −0.798 0.103 0.226 ns
Distance Temp.Path 35°C__neg 32°C__pos −0.141 −0.277 −0.005 0.038 *
Distance Temp.Path 35°C__pos 32°C__pos −0.449 −0.899 0.001 0.051 ns
Distance Temp.Path 35°C__pos 35°C__neg −0.308 −0.751 0.135 ≥0.25 ns
canberra
Distance Temp.Path 28°C__pos 28°C__neg 0.045 −0.074 0.164 ≥0.25 ns
Distance Temp.Path 32°C__neg 28°C__neg −0.123 −0.244 −0.002 0.045 *
Distance Temp.Path 32°C__pos 28°C__neg −0.042 −0.164 0.079 ≥0.25 ns
Distance Temp.Path 35°C__neg 28°C__neg −0.105 −0.214 0.005 0.068 ns
Distance Temp.Path 35°C__pos 28°C__neg −0.588 −0.883 −0.294 <0.001 ****
Distance Temp.Path 32°C__neg 28°C__pos −0.168 −0.267 −0.070 <0.001 ****
Distance Temp.Path 32°C__pos 28°C__pos −0.088 −0.186 0.010 0.107 ns
Distance Temp.Path 35°C__neg 28°C__pos −0.150 −0.233 −0.067 <0.001 ****
Distance Temp.Path 35°C__pos 28°C__pos −0.634 −0.919 −0.348 <0.001 ****
Distance Temp.Path 32°C__pos 32°C__neg 0.080 −0.021 0.182 0.199 ns
Distance Temp.Path 35°C__neg 32°C__neg 0.018 −0.068 0.105 ≥0.25 ns
Distance Temp.Path 35°C__pos 32°C__neg −0.465 −0.752 −0.179 <0.001 ***
Distance Temp.Path 35°C__neg 32°C__pos −0.062 −0.149 0.024 ≥0.25 ns
Distance Temp.Path 35°C__pos 32°C__pos −0.546 −0.832 −0.259 <0.001 ****
Distance Temp.Path 35°C__pos 35°C__neg −0.484 −0.765 −0.202 <0.001 ****
gunifrac
Distance Temp.Path 28°C__pos 28°C__neg 0.011 −0.129 0.151 ≥0.25 ns
Distance Temp.Path 32°C__neg 28°C__neg −0.091 −0.234 0.052 ≥0.25 ns
Distance Temp.Path 32°C__pos 28°C__neg −0.014 −0.157 0.129 ≥0.25 ns
Distance Temp.Path 35°C__neg 28°C__neg −0.129 −0.258 0.000 0.049 *
Distance Temp.Path 35°C__pos 28°C__neg −0.411 −0.758 −0.065 0.011 *
Distance Temp.Path 32°C__neg 28°C__pos −0.102 −0.218 0.014 0.115 ns
Distance Temp.Path 32°C__pos 28°C__pos −0.025 −0.141 0.091 ≥0.25 ns
Distance Temp.Path 35°C__neg 28°C__pos −0.140 −0.238 −0.043 <0.001 ***
Distance Temp.Path 35°C__pos 28°C__pos −0.423 −0.759 −0.087 0.006 **
Distance Temp.Path 32°C__pos 32°C__neg 0.077 −0.042 0.196 ≥0.25 ns
Distance Temp.Path 35°C__neg 32°C__neg −0.038 −0.140 0.064 ≥0.25 ns
Distance Temp.Path 35°C__pos 32°C__neg −0.321 −0.658 0.017 0.072 ns
Distance Temp.Path 35°C__neg 32°C__pos −0.115 −0.217 −0.013 0.017 *
Distance Temp.Path 35°C__pos 32°C__pos −0.398 −0.735 −0.060 0.011 *
Distance Temp.Path 35°C__pos 35°C__neg −0.283 −0.614 0.049 0.140 ns
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5C)

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GLM
GLM Results
glm(Total.Worm.Count ~ Alpha.Score*Cluster); Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) 0.000 0.778 0.000 ≥0.25 ns
Alpha.Score 0.000 1.808 0.000 ≥0.25 ns
ClusterLow 6.524 1.067 6.115 <0.001 ****
ClusterHigh −2.610 3.512 −0.743 ≥0.25 ns
Alpha.Score:ClusterLow −10.090 4.999 −2.018 0.047 *
Alpha.Score:ClusterHigh 13.706 4.766 2.876 0.005 **
Simpson
(Intercept) 0.000 0.741 0.000 ≥0.25 ns
Alpha.Score 0.000 1.605 0.000 ≥0.25 ns
ClusterLow 6.842 1.041 6.573 <0.001 ****
ClusterHigh −3.247 3.900 −0.833 ≥0.25 ns
Alpha.Score:ClusterLow −12.165 4.647 −2.618 0.011 *
Alpha.Score:ClusterHigh 14.198 5.090 2.789 0.007 **
Richness
(Intercept) 0.000 0.914 0.000 ≥0.25 ns
Alpha.Score 0.000 1.983 0.000 ≥0.25 ns
ClusterLow 6.105 1.294 4.717 <0.001 ****
ClusterHigh 7.362 2.644 2.784 0.007 **
Alpha.Score:ClusterLow −3.491 4.463 −0.782 ≥0.25 ns
Alpha.Score:ClusterHigh 0.772 4.163 0.185 ≥0.25 ns
Phylogenetic
(Intercept) 0.000 0.912 0.000 ≥0.25 ns
Alpha.Score 0.000 2.061 0.000 ≥0.25 ns
ClusterLow 6.230 1.346 4.629 <0.001 ****
ClusterHigh 8.137 2.589 3.143 0.002 **
Alpha.Score:ClusterLow −3.872 4.662 −0.831 ≥0.25 ns
Alpha.Score:ClusterHigh −0.429 4.237 −0.101 ≥0.25 ns
GLM Results
glm(Alpha.Score ~ Temperature*Total.Worm.Count); Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −1.421 0.364 −3.908 <0.001 ***
Temperature32 1.184 0.430 2.753 0.007 **
Temperature35 1.024 0.406 2.521 0.014 *
Total.Worm.Count 0.095 0.051 1.851 0.068 ns
Temperature32:Total.Worm.Count −0.030 0.066 −0.453 ≥0.25 ns
Temperature35:Total.Worm.Count −0.994 1.255 −0.792 ≥0.25 ns
Simpson
(Intercept) −1.579 0.386 −4.092 <0.001 ****
Temperature32 1.429 0.451 3.169 0.002 **
Temperature35 1.370 0.427 3.211 0.002 **
Total.Worm.Count 0.111 0.053 2.078 0.041 *
Temperature32:Total.Worm.Count −0.047 0.068 −0.687 ≥0.25 ns
Temperature35:Total.Worm.Count −0.889 1.182 −0.752 ≥0.25 ns
Richness
(Intercept) −0.749 0.284 −2.641 0.010 **
Temperature32 0.726 0.349 2.078 0.041 *
Temperature35 0.326 0.327 0.998 ≥0.25 ns
Total.Worm.Count 0.014 0.043 0.329 ≥0.25 ns
Temperature32:Total.Worm.Count 0.000 0.056 −0.006 ≥0.25 ns
Temperature35:Total.Worm.Count −0.625 1.049 −0.596 ≥0.25 ns
Phylogenetic
(Intercept) −0.705 0.267 −2.643 0.010 **
Temperature32 0.617 0.329 1.875 0.064 ns
Temperature35 0.199 0.308 0.645 ≥0.25 ns
Total.Worm.Count 0.005 0.041 0.123 ≥0.25 ns
Temperature32:Total.Worm.Count 0.012 0.053 0.221 ≥0.25 ns
Temperature35:Total.Worm.Count −0.507 0.986 −0.514 ≥0.25 ns
ANOVA
ANOVA of GLM
ANOVA(GLM(Total.Worm.Count ~ Alpha.Score*Cluster), type = 2); Exposed fish
term statistic df p.value sig
Shannon
Alpha.Score 0.143 1.000 ≥0.25 ns
Cluster 162.212 2.000 <0.001 ****
Alpha.Score:Cluster 14.204 2.000 <0.001 ***
Simpson
Alpha.Score 0.002 1.000 ≥0.25 ns
Cluster 168.513 2.000 <0.001 ****
Alpha.Score:Cluster 16.419 2.000 <0.001 ***
Richness
Alpha.Score 0.066 1.000 ≥0.25 ns
Cluster 145.539 2.000 <0.001 ****
Alpha.Score:Cluster 0.741 2.000 ≥0.25 ns
Phylogenetic
Alpha.Score 0.175 1.000 ≥0.25 ns
Cluster 145.520 2.000 <0.001 ****
Alpha.Score:Cluster 0.696 2.000 ≥0.25 ns
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Exposed fish
term statistic df p.value sig
Shannon
Temperature 13.111 2.000 0.001 **
Total.Worm.Count 5.701 1.000 0.017 *
Temperature:Total.Worm.Count 0.911 2.000 ≥0.25 ns
Simpson
Temperature 18.031 2.000 <0.001 ***
Total.Worm.Count 6.203 1.000 0.013 *
Temperature:Total.Worm.Count 1.061 2.000 ≥0.25 ns
Richness
Temperature 8.429 2.000 0.015 *
Total.Worm.Count 0.239 1.000 ≥0.25 ns
Temperature:Total.Worm.Count 0.386 2.000 ≥0.25 ns
Phylogenetic
Temperature 8.411 2.000 0.015 *
Total.Worm.Count 0.196 1.000 ≥0.25 ns
Temperature:Total.Worm.Count 0.339 2.000 ≥0.25 ns
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Total.Worm.Count ~ Alpha.Score*Cluster); Exposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score Cluster Low Other −1.594 0.642 −2.483 0.035 Cluster 28
28 Alpha.Score Cluster High Other 2.844 0.672 4.233 <0.001 Cluster 28
28 Alpha.Score Cluster High Low 4.438 0.773 5.741 <0.001 Cluster 28
32 Alpha.Score Cluster Low Other −1.391 0.495 −2.808 0.013 Cluster 32
32 Alpha.Score Cluster High Other 1.147 0.333 3.443 0.002 Cluster 32
32 Alpha.Score Cluster High Low 2.538 0.528 4.811 <0.001 Cluster 32
35 Alpha.Score Cluster Low Other −0.899 0.659 −1.365 0.172 Cluster 35
Simpson
28 Alpha.Score Cluster Low Other −1.487 0.656 −2.266 0.060 Cluster 28
28 Alpha.Score Cluster High Other 2.970 0.665 4.466 <0.001 Cluster 28
28 Alpha.Score Cluster High Low 4.457 0.767 5.810 <0.001 Cluster 28
32 Alpha.Score Cluster Low Other −1.233 0.487 −2.531 0.030 Cluster 32
32 Alpha.Score Cluster High Other 1.264 0.354 3.573 <0.001 Cluster 32
32 Alpha.Score Cluster High Low 2.497 0.528 4.729 <0.001 Cluster 32
35 Alpha.Score Cluster Low Other −0.779 0.692 −1.125 ≥0.25 Cluster 35
Richness
28 Alpha.Score Cluster Low Other −1.430 0.589 −2.429 0.040 Cluster 28
28 Alpha.Score Cluster High Other 1.367 0.653 2.095 0.090 Cluster 28
28 Alpha.Score Cluster High Low 2.797 0.692 4.043 <0.001 Cluster 28
32 Alpha.Score Cluster Low Other −1.183 0.387 −3.052 0.006 Cluster 32
32 Alpha.Score Cluster High Other 0.429 0.278 1.543 ≥0.25 Cluster 32
32 Alpha.Score Cluster High Low 1.611 0.410 3.934 <0.001 Cluster 32
35 Alpha.Score Cluster Low Other −0.611 0.552 −1.107 ≥0.25 Cluster 35
Phylogenetic
28 Alpha.Score Cluster Low Other −1.315 0.550 −2.389 0.044 Cluster 28
28 Alpha.Score Cluster High Other 1.204 0.611 1.970 0.119 Cluster 28
28 Alpha.Score Cluster High Low 2.519 0.638 3.946 <0.001 Cluster 28
32 Alpha.Score Cluster Low Other −0.994 0.360 −2.760 0.015 Cluster 32
32 Alpha.Score Cluster High Other 0.437 0.262 1.667 0.214 Cluster 32
32 Alpha.Score Cluster High Low 1.431 0.381 3.760 <0.001 Cluster 32
35 Alpha.Score Cluster Low Other −0.502 0.524 −0.958 ≥0.25 Cluster 35
Addl. Metrics

5D)

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ADONIS2
ADONIS2
adonis2(Beta Distance ~ Temperature*Cluster); Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 5.218 0.251 18.730 0.001 ***
Cluster 2.000 3.594 0.173 12.902 0.001 ***
Temperature:Cluster 3.000 0.653 0.031 1.564 0.097 ns
Residual 81.000 11.282 0.544 NA NA NA
Total 88.000 20.748 1.000 NA NA NA
canberra
Temperature 2.000 3.573 0.117 6.064 0.001 ***
Cluster 2.000 1.928 0.063 3.273 0.001 ***
Temperature:Cluster 3.000 1.157 0.038 1.310 0.052 ns
Residual 81.000 23.862 0.782 NA NA NA
Total 88.000 30.520 1.000 NA NA NA
gunifrac
Temperature 2.000 3.363 0.212 14.844 0.001 ***
Cluster 2.000 2.657 0.168 11.728 0.001 ***
Temperature:Cluster 3.000 0.645 0.041 1.898 0.027 *
Residual 81.000 9.176 0.579 NA NA NA
Total 88.000 15.841 1.000 NA NA NA
ADONIS2
adonis2(Beta Distance ~ Temperature*Total.Worm.Count); Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 5.218 0.251 14.815 0.001 ***
Total.Worm.Count 1.000 0.588 0.028 3.341 0.009 **
Temperature:Total.Worm.Count 2.000 0.326 0.016 0.925 ≥0.25 ns
Residual 83.000 14.616 0.704 NA NA NA
Total 88.000 20.748 1.000 NA NA NA
canberra
Temperature 2.000 3.573 0.117 5.754 0.001 ***
Total.Worm.Count 1.000 0.498 0.016 1.605 0.032 *
Temperature:Total.Worm.Count 2.000 0.678 0.022 1.093 ≥0.25 ns
Residual 83.000 25.771 0.844 NA NA NA
Total 88.000 30.520 1.000 NA NA NA
gunifrac
Temperature 2.000 3.363 0.212 11.736 0.001 ***
Total.Worm.Count 1.000 0.350 0.022 2.443 0.031 *
Temperature:Total.Worm.Count 2.000 0.235 0.015 0.822 ≥0.25 ns
Residual 83.000 11.892 0.751 NA NA NA
Total 88.000 15.841 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature*Cluster); Exposed fish
term df sumsq meansq statistic p.value sig
bray
Cluster 2.000 0.019 0.009 0.383 ≥0.25 ns
Residual 86.000 2.116 0.025 NA NA NA
canberra
Cluster 2.000 0.167 0.084 10.322 <0.001 ****
Residual 86.000 0.697 0.008 NA NA NA
gunifrac
Cluster 2.000 0.016 0.008 0.628 ≥0.25 ns
Residual 86.000 1.078 0.013 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Cluster); Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Cluster Low Other −0.037 −0.138 0.064 ≥0.25 ns
Distance Cluster High Other −0.005 −0.114 0.104 ≥0.25 ns
Distance Cluster High Low 0.032 −0.099 0.163 ≥0.25 ns
canberra
Distance Cluster Low Other 0.098 0.040 0.156 <0.001 ***
Distance Cluster High Other −0.031 −0.093 0.032 ≥0.25 ns
Distance Cluster High Low −0.129 −0.204 −0.053 <0.001 ***
gunifrac
Distance Cluster Low Other 0.017 −0.055 0.089 ≥0.25 ns
Distance Cluster High Other −0.027 −0.104 0.051 ≥0.25 ns
Distance Cluster High Low −0.044 −0.137 0.050 ≥0.25 ns
Addl. Metrics

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S5A

S5B

S5C

S5D

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Phylogenetic

ADONIS2
adonis2(Beta Distance ~ Cluster); Exposed fish
Beta.Metric term df SumOfSqs R2 statistic p.value p.adj.sig
Phylogenetic
bray Cluster 2.000 5.369 0.259 15.012 0.001 **
bray Residual 86.000 15.379 0.741 NA NA NA
bray Total 88.000 20.748 1.000 NA NA NA
canberra Cluster 2.000 3.204 0.105 5.043 0.001 **
canberra Residual 86.000 27.317 0.895 NA NA NA
canberra Total 88.000 30.520 1.000 NA NA NA
gunifrac Cluster 2.000 3.692 0.233 13.068 0.001 **
gunifrac Residual 86.000 12.149 0.767 NA NA NA
gunifrac Total 88.000 15.841 1.000 NA NA NA
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Dispersion ~ Cluster); Exposed fish
Beta.Metric term df sumsq meansq statistic p.value p.adj.sig
Phylogenetic
bray Groups 2.000 0.039 0.019 0.754 ≥0.25 ns
bray Residuals 86.000 2.218 0.026 NA NA NA
canberra Groups 2.000 0.221 0.110 14.247 <0.001 ****
canberra Residuals 86.000 0.666 0.008 NA NA NA
gunifrac Groups 2.000 0.072 0.036 2.725 0.071 ns
gunifrac Residuals 86.000 1.131 0.013 NA NA NA
Richness

ADONIS2
adonis2(Beta Distance ~ Cluster); Exposed fish
Beta.Metric term df SumOfSqs R2 statistic p.value p.adj.sig
Richness
bray Cluster 2.000 5.363 0.258 14.990 0.001 **
bray Residual 86.000 15.385 0.742 NA NA NA
bray Total 88.000 20.748 1.000 NA NA NA
canberra Cluster 2.000 3.356 0.110 5.312 0.001 **
canberra Residual 86.000 27.165 0.890 NA NA NA
canberra Total 88.000 30.520 1.000 NA NA NA
gunifrac Cluster 2.000 3.835 0.242 13.736 0.001 **
gunifrac Residual 86.000 12.006 0.758 NA NA NA
gunifrac Total 88.000 15.841 1.000 NA NA NA
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Dispersion ~ Cluster); Exposed fish
Beta.Metric term df sumsq meansq statistic p.value p.adj.sig
Richness
bray Groups 2.000 0.028 0.014 0.535 ≥0.25 ns
bray Residuals 86.000 2.263 0.026 NA NA NA
canberra Groups 2.000 0.216 0.108 13.957 <0.001 ****
canberra Residuals 86.000 0.664 0.008 NA NA NA
gunifrac Groups 2.000 0.050 0.025 1.873 0.160 ns
gunifrac Residuals 86.000 1.155 0.013 NA NA NA
Shannon

ADONIS2
adonis2(Beta Distance ~ Cluster); Exposed fish
Beta.Metric term df SumOfSqs R2 statistic p.value p.adj.sig
Shannon
bray Cluster 2.000 5.943 0.286 17.260 0.001 **
bray Residual 86.000 14.805 0.714 NA NA NA
bray Total 88.000 20.748 1.000 NA NA NA
canberra Cluster 2.000 3.188 0.104 5.016 0.001 **
canberra Residual 86.000 27.332 0.896 NA NA NA
canberra Total 88.000 30.520 1.000 NA NA NA
gunifrac Cluster 2.000 4.229 0.267 15.660 0.001 **
gunifrac Residual 86.000 11.612 0.733 NA NA NA
gunifrac Total 88.000 15.841 1.000 NA NA NA
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Dispersion ~ Cluster); Exposed fish
Beta.Metric term df sumsq meansq statistic p.value p.adj.sig
Shannon
bray Groups 2.000 0.019 0.009 0.383 ≥0.25 ns
bray Residuals 86.000 2.116 0.025 NA NA NA
canberra Groups 2.000 0.167 0.084 10.322 <0.001 ****
canberra Residuals 86.000 0.697 0.008 NA NA NA
gunifrac Groups 2.000 0.016 0.008 0.628 ≥0.25 ns
gunifrac Residuals 86.000 1.078 0.013 NA NA NA
Simpsons

ADONIS2
adonis2(Beta Distance ~ Cluster); Exposed fish
Beta.Metric term df SumOfSqs R2 statistic p.value p.adj.sig
Simpson
bray Cluster 2.000 5.943 0.286 17.260 0.001 **
bray Residual 86.000 14.805 0.714 NA NA NA
bray Total 88.000 20.748 1.000 NA NA NA
canberra Cluster 2.000 3.188 0.104 5.016 0.001 **
canberra Residual 86.000 27.332 0.896 NA NA NA
canberra Total 88.000 30.520 1.000 NA NA NA
gunifrac Cluster 2.000 4.229 0.267 15.660 0.001 **
gunifrac Residual 86.000 11.612 0.733 NA NA NA
gunifrac Total 88.000 15.841 1.000 NA NA NA
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Dispersion ~ Cluster); Exposed fish
Beta.Metric term df sumsq meansq statistic p.value p.adj.sig
Simpson
bray Groups 2.000 0.019 0.009 0.383 ≥0.25 ns
bray Residuals 86.000 2.116 0.025 NA NA NA
canberra Groups 2.000 0.167 0.084 10.322 <0.001 ****
canberra Residuals 86.000 0.697 0.008 NA NA NA
gunifrac Groups 2.000 0.016 0.008 0.628 ≥0.25 ns
gunifrac Residuals 86.000 1.078 0.013 NA NA NA

Cluster Counts

Counts of samples per cluster and temperature by alpha-diversity metric. Click on tabs to reveal tables.

(hide)

Simpson

Temperature Count Percentage
Simpson - High
28 5 33.33
32 10 66.67
Simpson - Low
28 12 66.67
32 5 27.78
35 1 5.56
Simpson - Other
28 8 14.29
32 15 26.79
35 33 58.93

Shannon

Temperature Count Percentage
Shannon - High
28 5 33.33
32 10 66.67
Shannon - Low
28 12 66.67
32 5 27.78
35 1 5.56
Shannon - Other
28 8 14.29
32 15 26.79
35 33 58.93

Richness

Temperature Count Percentage
Richness - High
28 5 35.71
32 9 64.29
Richness - Low
28 12 63.16
32 6 31.58
35 1 5.26
Richness - Other
28 8 14.29
32 15 26.79
35 33 58.93

Phylogenetic

Temperature Count Percentage
Phylogenetic - High
28 4 30.77
32 9 69.23
Phylogenetic - Low
28 13 65.00
32 6 30.00
35 1 5.00
Phylogenetic - Other
28 8 14.29
32 15 26.79
35 33 58.93

Fig. 6) Parasite exposure exacerbates water temperature differences in gut microbiome structure

Fig. 6 Comparison of the effects of water temperature on the gut microbiome between parasite exposed fish and parasite unexposed fish. (A) Simpson’s Index for diversity of parasite unexposed and pre-exposed fish at 0 days post exposure (dpe). Prior to parasite exposure gut microbial alpha-diversity does not significantly differ between fish reared at the same water temperature. Capscale ordinations based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main and interaction effects of temperature and parasite exposure (treatment) (B) of pre-exposure samples at 0 dpe. (C) Simpson’s Index for diversity of parasite unexposed and exposed fish. Gut microbial alpha-diversity significantly differs between parasite exposed fish reared at 28°C and 32°C water temperature relative to unexposed control fish, but gut microbial alpha-diversity does not differ between parasite unexposed and exposed fish reared at 35°C water temperature. Capscale ordinations based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main and interaction effects of temperature and parasite exposure (treatment) of (D) post-exposure samples after 0 dpe. The analysis shows gut microbiome composition differs between fish reared at different water temperatures prior to parasite exposure, and parasite exposure further drives these temperature associated differences in microbiome community composition. Ribbons and ellipses indicate 95% confidence interval. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate statistically significant covariates and direction of greatest change in the indicated covariates.

6A)

Tables
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GLM
GLM Results
glm(Alpha.Score ~ Temperature*Treatment); Pre-Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −0.218 0.269 −0.811 ≥0.25 ns
Temperature32 0.345 0.379 0.910 ≥0.25 ns
Temperature35 0.693 0.384 1.803 0.077 ns
TreatmentExposed 0.274 0.379 0.723 ≥0.25 ns
Temperature32:TreatmentExposed 0.311 0.544 0.572 ≥0.25 ns
Temperature35:TreatmentExposed 0.041 0.550 0.075 ≥0.25 ns
Simpson
(Intercept) −0.241 0.252 −0.958 ≥0.25 ns
Temperature32 0.352 0.355 0.990 ≥0.25 ns
Temperature35 0.961 0.367 2.619 0.011 *
TreatmentExposed 0.223 0.355 0.627 ≥0.25 ns
Temperature32:TreatmentExposed 0.486 0.513 0.948 ≥0.25 ns
Temperature35:TreatmentExposed 0.017 0.525 0.033 ≥0.25 ns
Richness
(Intercept) −0.145 0.270 −0.538 ≥0.25 ns
Temperature32 0.550 0.385 1.430 0.158 ns
Temperature35 0.043 0.381 0.112 ≥0.25 ns
TreatmentExposed 0.402 0.382 1.050 ≥0.25 ns
Temperature32:TreatmentExposed 0.020 0.554 0.036 ≥0.25 ns
Temperature35:TreatmentExposed 0.287 0.545 0.526 ≥0.25 ns
Phylogenetic
(Intercept) −0.253 0.254 −0.996 ≥0.25 ns
Temperature32 0.553 0.360 1.536 0.130 ns
Temperature35 0.171 0.358 0.478 ≥0.25 ns
TreatmentExposed 0.335 0.358 0.937 ≥0.25 ns
Temperature32:TreatmentExposed −0.018 0.512 −0.035 ≥0.25 ns
Temperature35:TreatmentExposed 0.102 0.507 0.201 ≥0.25 ns
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Treatment), type = 2); Pre-Exposed fish
term statistic df p.value sig
Shannon
Temperature 7.264 2.000 0.026 *
Treatment 3.044 1.000 0.081 ns
Temperature:Treatment 0.380 2.000 ≥0.25 ns
Simpson
Temperature 14.462 2.000 <0.001 ***
Treatment 3.406 1.000 0.065 ns
Temperature:Treatment 1.122 2.000 ≥0.25 ns
Richness
Temperature 4.311 2.000 0.116 ns
Treatment 5.067 1.000 0.024 *
Temperature:Treatment 0.340 2.000 ≥0.25 ns
Phylogenetic
Temperature 4.600 2.000 0.100 ns
Treatment 3.056 1.000 0.080 ns
Temperature:Treatment 0.064 2.000 ≥0.25 ns
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Treatment); Pre-Exposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score Treatment Exposed Control 0.274 0.364 0.753 ≥0.25 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.585 0.451 1.298 0.194 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.315 0.344 0.917 ≥0.25 Treatment 35
Simpson
28 Alpha.Score Treatment Exposed Control 0.223 0.337 0.662 ≥0.25 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.709 0.426 1.663 0.096 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.240 0.339 0.708 ≥0.25 Treatment 35
Richness
28 Alpha.Score Treatment Exposed Control 0.402 0.361 1.112 ≥0.25 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.422 0.411 1.027 ≥0.25 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.689 0.400 1.721 0.085 Treatment 35
Phylogenetic
28 Alpha.Score Treatment Exposed Control 0.335 0.344 0.974 ≥0.25 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.317 0.348 0.911 ≥0.25 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.437 0.389 1.123 ≥0.25 Treatment 35
Addl. Metrics

6B)

Tables
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ADONIS2
ADONIS2
adonis2(Beta Distance ~ Temperature*Treatment); Pre-Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 0.871 0.188 7.178 0.001 ***
Treatment 1.000 0.244 0.053 4.025 0.009 **
Temperature:Treatment 2.000 0.245 0.053 2.019 0.047 *
Residual 54.000 3.275 0.707 NA NA NA
Total 59.000 4.635 1.000 NA NA NA
canberra
Temperature 2.000 1.652 0.132 4.615 0.001 ***
Treatment 1.000 0.533 0.043 2.979 0.001 ***
Temperature:Treatment 2.000 0.667 0.053 1.864 0.001 ***
Residual 54.000 9.665 0.772 NA NA NA
Total 59.000 12.517 1.000 NA NA NA
gunifrac
Temperature 2.000 0.647 0.153 5.604 0.001 ***
Treatment 1.000 0.288 0.068 4.987 0.003 **
Temperature:Treatment 2.000 0.177 0.042 1.537 0.110 ns
Residual 54.000 3.117 0.737 NA NA NA
Total 59.000 4.229 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Pre-Exposed fish
term df sumsq meansq statistic p.value sig
bray
Temp.Treat 5.000 0.103 0.021 2.738 0.028 *
Residual 54.000 0.405 0.007 NA NA NA
canberra
Temp.Treat 5.000 0.014 0.003 0.453 ≥0.25 ns
Residual 54.000 0.329 0.006 NA NA NA
gunifrac
Temp.Treat 5.000 0.054 0.011 1.564 0.186 ns
Residual 54.000 0.369 0.007 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Treatment); Pre-Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.027 −0.088 0.141 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Control 0.095 −0.019 0.210 0.153 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.110 −0.004 0.225 0.065 ns
Distance Temp.Treat 35°C__Control 28°C__Control 0.015 −0.099 0.130 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.069 −0.045 0.183 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Exposed 0.069 −0.046 0.183 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Exposed 0.084 −0.031 0.198 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.011 −0.126 0.103 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Exposed 0.042 −0.072 0.157 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.015 −0.100 0.129 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Control −0.080 −0.195 0.034 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control −0.026 −0.141 0.088 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.095 −0.209 0.019 0.156 ns
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.041 −0.156 0.073 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.054 −0.061 0.168 ≥0.25 ns
canberra
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.011 −0.092 0.114 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Control 0.035 −0.068 0.138 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.044 −0.059 0.147 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Control 0.010 −0.094 0.113 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.016 −0.087 0.119 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Exposed 0.024 −0.079 0.127 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Exposed 0.033 −0.070 0.136 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.001 −0.104 0.102 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Exposed 0.005 −0.098 0.108 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.009 −0.094 0.112 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Control −0.025 −0.128 0.078 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control −0.019 −0.122 0.084 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.034 −0.137 0.069 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.028 −0.131 0.075 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.006 −0.097 0.109 ≥0.25 ns
gunifrac
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.003 −0.106 0.112 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Control 0.051 −0.058 0.160 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.078 −0.032 0.187 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Control 0.000 −0.109 0.110 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.042 −0.067 0.151 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Exposed 0.048 −0.061 0.158 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Exposed 0.075 −0.035 0.184 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.002 −0.112 0.107 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Exposed 0.039 −0.070 0.148 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.026 −0.083 0.136 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Control −0.051 −0.160 0.059 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control −0.009 −0.118 0.100 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.077 −0.186 0.032 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.036 −0.145 0.074 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.041 −0.068 0.151 ≥0.25 ns
Addl. Metrics

6C)

Tables
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GLM
GLM Results
glm(Alpha.Score ~ Temperature*Treatment); Post-Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −0.441 0.126 −3.499 <0.001 ***
Temperature32 −0.064 0.177 −0.364 ≥0.25 ns
Temperature35 −0.087 0.182 −0.476 ≥0.25 ns
TreatmentExposed −0.539 0.210 −2.562 0.011 *
Temperature32:TreatmentExposed 0.953 0.280 3.405 <0.001 ***
Temperature35:TreatmentExposed 0.561 0.281 1.996 0.047 *
Simpson
(Intercept) −0.299 0.133 −2.252 0.025 *
Temperature32 −0.059 0.186 −0.319 ≥0.25 ns
Temperature35 0.080 0.190 0.419 ≥0.25 ns
TreatmentExposed −0.724 0.225 −3.223 0.001 **
Temperature32:TreatmentExposed 1.106 0.298 3.715 <0.001 ***
Temperature35:TreatmentExposed 0.673 0.297 2.270 0.024 *
Richness
(Intercept) −0.206 0.101 −2.040 0.043 *
Temperature32 −0.048 0.141 −0.343 ≥0.25 ns
Temperature35 −0.317 0.147 −2.164 0.032 *
TreatmentExposed −0.228 0.160 −1.421 0.157 ns
Temperature32:TreatmentExposed 0.829 0.219 3.776 <0.001 ***
Temperature35:TreatmentExposed 0.691 0.219 3.155 0.002 **
Phylogenetic
(Intercept) −0.143 0.095 −1.507 0.134 ns
Temperature32 −0.035 0.132 −0.268 ≥0.25 ns
Temperature35 −0.319 0.138 −2.320 0.021 *
TreatmentExposed −0.192 0.150 −1.280 0.202 ns
Temperature32:TreatmentExposed 0.780 0.206 3.783 <0.001 ***
Temperature35:TreatmentExposed 0.639 0.205 3.111 0.002 **
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Treatment), type = 2); Post-Exposed fish
term statistic df p.value sig
Shannon
Temperature 5.908 2.000 0.052 ns
Treatment 0.001 1.000 ≥0.25 ns
Temperature:Treatment 11.784 2.000 0.003 **
Simpson
Temperature 8.400 2.000 0.015 *
Treatment 0.547 1.000 ≥0.25 ns
Temperature:Treatment 14.128 2.000 <0.001 ***
Richness
Temperature 11.325 2.000 0.003 **
Treatment 11.824 1.000 <0.001 ***
Temperature:Treatment 16.178 2.000 <0.001 ***
Phylogenetic
Temperature 13.345 2.000 0.001 **
Treatment 13.294 1.000 <0.001 ***
Temperature:Treatment 16.080 2.000 <0.001 ***
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Treatment); Post-Exposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score Treatment Exposed Control −0.539 0.280 −1.924 0.054 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.414 0.180 2.298 0.022 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.022 0.111 0.200 ≥0.25 Treatment 35
Simpson
28 Alpha.Score Treatment Exposed Control −0.724 0.290 −2.492 0.013 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.382 0.190 2.013 0.044 Treatment 32
35 Alpha.Score Treatment Exposed Control −0.051 0.130 −0.387 ≥0.25 Treatment 35
Richness
28 Alpha.Score Treatment Exposed Control −0.228 0.223 −1.021 ≥0.25 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.601 0.132 4.546 <0.001 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.463 0.092 5.021 <0.001 Treatment 35
Phylogenetic
28 Alpha.Score Treatment Exposed Control −0.192 0.210 −0.916 ≥0.25 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.588 0.121 4.858 <0.001 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.447 0.090 4.961 <0.001 Treatment 35
Addl. Metrics

6D)

Tables
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ADONIS2
ADONIS2
adonis2(Beta Distance ~ Temperature*Treatment); Post-Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 4.123 0.108 17.035 0.001 ***
Treatment 1.000 7.411 0.194 61.235 0.001 ***
Temperature:Treatment 2.000 3.168 0.083 13.088 0.001 ***
Residual 194.000 23.478 0.615 NA NA NA
Total 199.000 38.179 1.000 NA NA NA
canberra
Temperature 2.000 4.561 0.074 9.502 0.001 ***
Treatment 1.000 7.402 0.120 30.840 0.001 ***
Temperature:Treatment 2.000 3.014 0.049 6.279 0.001 ***
Residual 194.000 46.561 0.757 NA NA NA
Total 199.000 61.538 1.000 NA NA NA
gunifrac
Temperature 2.000 2.273 0.082 11.945 0.001 ***
Treatment 1.000 4.718 0.170 49.577 0.001 ***
Temperature:Treatment 2.000 2.273 0.082 11.944 0.001 ***
Residual 194.000 18.460 0.666 NA NA NA
Total 199.000 27.724 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Post-Exposed fish
term df sumsq meansq statistic p.value sig
bray
Temp.Treat 5.000 1.344 0.269 17.882 <0.001 ****
Residual 194.000 2.917 0.015 NA NA NA
canberra
Temp.Treat 5.000 1.094 0.219 31.758 <0.001 ****
Residual 194.000 1.337 0.007 NA NA NA
gunifrac
Temp.Treat 5.000 1.225 0.245 29.264 <0.001 ****
Residual 194.000 1.624 0.008 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Treatment); Post-Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.211 0.119 0.302 <0.001 ****
Distance Temp.Treat 32°C__Control 28°C__Control −0.015 −0.096 0.066 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.168 0.081 0.255 <0.001 ****
Distance Temp.Treat 35°C__Control 28°C__Control 0.013 −0.070 0.096 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.060 −0.024 0.144 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Exposed −0.225 −0.316 −0.135 <0.001 ****
Distance Temp.Treat 32°C__Exposed 28°C__Exposed −0.043 −0.138 0.053 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.197 −0.290 −0.105 <0.001 ****
Distance Temp.Treat 35°C__Exposed 28°C__Exposed −0.150 −0.243 −0.057 <0.001 ****
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.183 0.097 0.268 <0.001 ****
Distance Temp.Treat 35°C__Control 32°C__Control 0.028 −0.054 0.110 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control 0.075 −0.008 0.158 0.100 ns
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.155 −0.243 −0.067 <0.001 ****
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.108 −0.196 −0.019 0.007 **
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.047 −0.038 0.132 ≥0.25 ns
canberra
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.217 0.155 0.279 <0.001 ****
Distance Temp.Treat 32°C__Control 28°C__Control −0.011 −0.066 0.044 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.107 0.048 0.166 <0.001 ****
Distance Temp.Treat 35°C__Control 28°C__Control 0.013 −0.044 0.069 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.073 0.016 0.130 0.004 **
Distance Temp.Treat 32°C__Control 28°C__Exposed −0.228 −0.289 −0.167 <0.001 ****
Distance Temp.Treat 32°C__Exposed 28°C__Exposed −0.110 −0.175 −0.045 <0.001 ****
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.204 −0.267 −0.142 <0.001 ****
Distance Temp.Treat 35°C__Exposed 28°C__Exposed −0.144 −0.207 −0.081 <0.001 ****
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.118 0.060 0.176 <0.001 ****
Distance Temp.Treat 35°C__Control 32°C__Control 0.024 −0.032 0.080 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control 0.084 0.028 0.140 <0.001 ***
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.094 −0.154 −0.035 <0.001 ***
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.034 −0.094 0.026 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.060 0.003 0.118 0.035 *
gunifrac
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.197 0.129 0.266 <0.001 ****
Distance Temp.Treat 32°C__Control 28°C__Control −0.023 −0.083 0.037 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.141 0.077 0.206 <0.001 ****
Distance Temp.Treat 35°C__Control 28°C__Control −0.011 −0.073 0.051 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.054 −0.009 0.117 0.134 ns
Distance Temp.Treat 32°C__Control 28°C__Exposed −0.220 −0.288 −0.153 <0.001 ****
Distance Temp.Treat 32°C__Exposed 28°C__Exposed −0.056 −0.128 0.015 0.211 ns
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.209 −0.278 −0.140 <0.001 ****
Distance Temp.Treat 35°C__Exposed 28°C__Exposed −0.143 −0.213 −0.074 <0.001 ****
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.164 0.100 0.228 <0.001 ****
Distance Temp.Treat 35°C__Control 32°C__Control 0.012 −0.050 0.073 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control 0.077 0.015 0.139 0.006 **
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.152 −0.218 −0.087 <0.001 ****
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.087 −0.153 −0.021 0.003 **
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.065 0.002 0.129 0.039 *
Addl. Metrics

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S6A

S6B

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By Temp
## NULL
## NULL
## NULL
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ADONIS2
Temperature term df SumOfSqs R2 statistic p.value p.adj.sig
bray
28 Treatment 1.000 0.065 0.082 1.615 0.176 ns
28 Residual 18.000 0.723 0.918 NA NA NA
28 Total 19.000 0.788 1.000 NA NA NA
32 Treatment 1.000 0.195 0.109 2.191 0.087 ns
32 Residual 18.000 1.603 0.891 NA NA NA
32 Total 19.000 1.799 1.000 NA NA NA
35 Treatment 1.000 0.229 0.194 4.346 0.008 **
35 Residual 18.000 0.948 0.806 NA NA NA
35 Total 19.000 1.177 1.000 NA NA NA
canberra
28 Treatment 1.000 0.358 0.105 2.102 0.010 *
28 Residual 18.000 3.068 0.895 NA NA NA
28 Total 19.000 3.426 1.000 NA NA NA
32 Treatment 1.000 0.422 0.107 2.153 0.009 **
32 Residual 18.000 3.524 0.893 NA NA NA
32 Total 19.000 3.946 1.000 NA NA NA
35 Treatment 1.000 0.421 0.120 2.464 0.007 **
35 Residual 18.000 3.073 0.880 NA NA NA
35 Total 19.000 3.493 1.000 NA NA NA
gunifrac
28 Treatment 1.000 0.086 0.092 1.828 0.130 ns
28 Residual 18.000 0.845 0.908 NA NA NA
28 Total 19.000 0.931 1.000 NA NA NA
32 Treatment 1.000 0.174 0.113 2.296 0.062 ns
32 Residual 18.000 1.361 0.887 NA NA NA
32 Total 19.000 1.535 1.000 NA NA NA
35 Treatment 1.000 0.206 0.184 4.068 0.012 *
35 Residual 18.000 0.911 0.816 NA NA NA
35 Total 19.000 1.117 1.000 NA NA NA
Dispersion (ANOVA)
Temperature term df sumsq meansq statistic p.value p.adj.sig
bray
28 Groups 1.000 0.004 0.004 0.609 0.445 ns
28 Residuals 18.000 0.105 0.006 NA NA NA
32 Groups 1.000 0.001 0.001 0.100 0.755 ns
32 Residuals 18.000 0.199 0.011 NA NA NA
35 Groups 1.000 0.014 0.014 2.589 0.125 ns
35 Residuals 18.000 0.101 0.006 NA NA NA
canberra
28 Groups 1.000 0.001 0.001 0.061 0.807 ns
28 Residuals 18.000 0.172 0.010 NA NA NA
32 Groups 1.000 0.000 0.000 0.078 0.783 ns
32 Residuals 18.000 0.092 0.005 NA NA NA
35 Groups 1.000 0.000 0.000 0.055 0.817 ns
35 Residuals 18.000 0.065 0.004 NA NA NA
gunifrac
28 Groups 1.000 0.000 0.000 0.004 0.948 ns
28 Residuals 18.000 0.165 0.009 NA NA NA
32 Groups 1.000 0.003 0.003 0.465 0.504 ns
32 Residuals 18.000 0.135 0.008 NA NA NA
35 Groups 1.000 0.009 0.009 2.238 0.152 ns
35 Residuals 18.000 0.069 0.004 NA NA NA

S6B

S6D

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ADONIS2
Temperature term df SumOfSqs R2 statistic p.value p.adj.sig
bray
28 Treatment 1.000 6.719 0.447 48.541 0.001 **
28 Residual 60.000 8.305 0.553 NA NA NA
28 Total 61.000 15.023 1.000 NA NA NA
32 Treatment 1.000 2.842 0.261 23.668 0.001 **
32 Residual 67.000 8.044 0.739 NA NA NA
32 Total 68.000 10.886 1.000 NA NA NA
35 Treatment 1.000 1.018 0.125 9.568 0.001 **
35 Residual 67.000 7.129 0.875 NA NA NA
35 Total 68.000 8.147 1.000 NA NA NA
canberra
28 Treatment 1.000 3.887 0.191 14.144 0.001 **
28 Residual 60.000 16.490 0.809 NA NA NA
28 Total 61.000 20.377 1.000 NA NA NA
32 Treatment 1.000 3.542 0.191 15.809 0.001 **
32 Residual 67.000 15.012 0.809 NA NA NA
32 Total 68.000 18.554 1.000 NA NA NA
35 Treatment 1.000 2.987 0.165 13.288 0.001 **
35 Residual 67.000 15.059 0.835 NA NA NA
35 Total 68.000 18.046 1.000 NA NA NA
gunifrac
28 Treatment 1.000 4.085 0.374 35.857 0.001 **
28 Residual 60.000 6.836 0.626 NA NA NA
28 Total 61.000 10.922 1.000 NA NA NA
32 Treatment 1.000 2.031 0.230 19.988 0.001 **
32 Residual 67.000 6.806 0.770 NA NA NA
32 Total 68.000 8.837 1.000 NA NA NA
35 Treatment 1.000 1.080 0.160 12.720 0.001 **
35 Residual 67.000 5.691 0.840 NA NA NA
35 Total 68.000 6.771 1.000 NA NA NA
Dispersion (ANOVA)
Temperature term df sumsq meansq statistic p.value p.adj.sig
bray
28 Groups 1.000 0.662 0.662 53.570 0.000 ****
28 Residuals 60.000 0.741 0.012 NA NA NA
32 Groups 1.000 0.567 0.567 54.435 0.000 ****
32 Residuals 67.000 0.697 0.010 NA NA NA
35 Groups 1.000 0.038 0.038 1.735 0.192 ns
35 Residuals 67.000 1.479 0.022 NA NA NA
canberra
28 Groups 1.000 0.703 0.703 224.734 0.000 ****
28 Residuals 60.000 0.188 0.003 NA NA NA
32 Groups 1.000 0.237 0.237 30.033 0.000 ****
32 Residuals 67.000 0.530 0.008 NA NA NA
35 Groups 1.000 0.062 0.062 6.749 0.012 *
35 Residuals 67.000 0.620 0.009 NA NA NA
gunifrac
28 Groups 1.000 0.582 0.582 124.383 0.000 ****
28 Residuals 60.000 0.281 0.005 NA NA NA
32 Groups 1.000 0.482 0.482 57.322 0.000 ****
32 Residuals 67.000 0.564 0.008 NA NA NA
35 Groups 1.000 0.078 0.078 5.842 0.018 *
35 Residuals 67.000 0.896 0.013 NA NA NA

Fig. 7) Gut microbial abundance is significantly associated with environmental conditions and stressors

7) Gut microbial abundance is significantly associated with environmental conditions and stressors

Fig. 7 A heatmap of model coefficient values of the top 50 statistically significant abundant gut microbial taxa identified by MaAsLin2. The color of each cell represents the coefficient value and direction (red is positive, blue is negative). A “+” or “-” indicates a statistically significant association was observed between taxon abundance and a covariate. Gray colored cells indicate a significant effect was not observed.

Taxon Temp: 32°C Temp: 35°C Time (DPE) Cluster: Low Parasite exposed Infection present Infection burden
Actinobacteriota
IMCC26207 +
- - -

Nocardiaceae Genus + + +



PeM15 Genus +
- - -

Bacteroidota
Barnesiellaceae Genus

-
-

Cloacibacterium
+ -



Flavobacterium - - - - +

Fluviicola
- -
- -
Microscillaceae Genus + + + -


Terrimonas


-
+ +
env OPS 17 Genus
-
- -

Chloroflexi
JG30 KF CM45 Genus

+ - + +
Cyanobacteria
Candidatus Obscuribacter - - + - + +
Obscuribacteraceae Genus +
+ -
+ -
Vampirovibrionaceae Genus

+ - + +
Desulfobacterota
Desulfobacterota Genus


- + +
Firmicutes
Clostridium sensu stricto 3

+ - + +
Gemmatimonadota
Gemmatimonadaceae Genus
-
- + +
Myxococcota
Sandaracinaceae Genus + + + - + +
NB1-j
NB1 j Genus
+ + - + +
Nitrospirota
Nitrospira
- + - + +
Patescibacteria
Saccharimonadales Genus + + + - +

Planctomycetota
CL500 3

+ - - +
Gemmata
+ + - + +
Pirellulaceae Genus + - + - -

Planctomycetales Genus

+ - + +
Planctomycetes Genus

+ - + +
Rhodopirellula


- + + -
SM1A02

+ - + +
Telmatocola

+ - + +
Proteobacteria
A0839 Genus +

- + +
Alphaproteobacteria Genus
+ + - + +
Bauldia +
+ - + +
Beijerinckiaceae Genus
- + -
+ -
Bosea - - - -


Candidatus Alysiosphaera +
+ - + +
Chitinibacter
+ +
+
-
Comamonadaceae Genus

- - + +
Gammaproteobacteria Genus - - + -
+
Luteimonas -

-
+ +
Methyloligellaceae Genus + + + - + +
Nordella
-
- + +
Plesiomonas + +

- - -
Pseudomonadales Genus - - - - - -
Rhizobacter


- + + +
Rhizobiales Incertae Sedis Genus


-
+
Rhodobacteraceae Genus


- + +
Rhodovastum
-
-


Steroidobacteraceae Genus

+ - + +
Xanthobacteraceae Genus - - + -
+
Verrucomicrobiota
Oikopleura

+ - + +
Tables
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S7.1
Summary of Significant Associations
(per covariate)
Significance Cluster (Low) DPE (DPE) Pathology.Results (positive) Temperature (32) Temperature (35) Total.Worm.Count (Total.Worm.Count) Treatment (Exposed)
Positive 1 73 117 42 37 49 74
Negative 192 36 5 47 54 13 35
S7.2
Summary of Significant Associations
(per taxonomic level)
Taxonomy Count
Kingdom 3
Phylum 28
Class 48
Order 114
Family 183
Genus 277
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S7.1
All taxa with significant associations
MaAsLin2(log(qval)*sign(coef)); All fish
Taxon Temp: 32°C Temp: 35°C Time (DPE) Cluster: Low Parasite exposed Infection present Infection burden
Acidobacteriota
Bryobacter





+
Holophagaceae Genus

- -
+
JGI 0001001 H03


-
+ +
Paludibaculum Genus +

- +

Subgroup 17 Genus


- +

Vicinamibacteraceae Genus


-
+ +
Actinobacteriota
Acidimicrobiia Genus
+

-

Agromyces
+

+

Brevibacterium

+



Conexibacter
+ +
+

Corynebacteriales Genus - - + -


Frankiales Genus

+ - + +
Gaiella


-

+
Gaiellales Genus +
+ -


Glutamicibacter


-


Gordonia + + + -


IMCC26207 +
- - -

IMCC26256 Genus +
+ -
+
Iamia
- + -


Ilumatobacteraceae Genus

+
+

Kineosporiaceae Genus

+



Leucobacter

-



Longivirga
+
-


Microtrichaceae Genus





+
Microtrichales Genus + + - -


Mycobacterium -
+ - - +
Nakamurella


-


Nocardia


-
+
Nocardiaceae Genus + + +



Nocardioides

+ -


PeM15 Genus +
- - -

Rhodococcus + + +
-

Smaragdicoccus + + +



Thermoleophilia Genus


- +
+
Armatimonadota
Armatimonadota Genus





+
Bacteroidota
AKYH767 Genus -

-
+
Aurantisolimonas

-
-

Bacteroidia Genus -
- -
+
Barnesiellaceae Genus

-
-

Candidatus Amoebophilus

+



Chitinophagaceae Genus -

-
+ +
Chitinophagales Genus -

-
+
Chryseobacterium
+




Cloacibacterium
+ -



Cytophaga -


+

Cytophagales Genus -

-
+
Dinghuibacter + +
- -

Ferruginibacter - -
-
+ +
Flavihumibacter
- - - -

Flavobacterium - - - - +

Fluviicola
- -
- -
Haliscomenobacter - -
-
+
Hassallia


-

+
Kapabacteriales Genus -

-
+
Microscillaceae Genus + + + -


NS9 marine group Genus

+ - +

OLB12 +
+ - +

Ohtaekwangia
+ +
+

Runella -

-
+
Saprospiraceae Genus -

-

+
Sediminibacterium -





Sphingobacteriales Genus


-
+
Terrimonas


-
+ +
env OPS 17 Genus
-
- -

Bdellovibrionota
Bacteriovoracaceae Genus -

-
+
Bdellovibrio
-
-
+
OM27 clade


-
+ +
Oligoflexaceae Genus
-




Chloroflexi
Caldilineaceae Genus


-
+ +
JG30 KF CM45 Genus

+ - + +
KD4 96 Genus

+ -
+
RBG 13 54 9 Genus -

-
+ +
SBR1031 Genus


-


Cyanobacteria
Candidatus Obscuribacter - - + - + +
Obscuribacteraceae Genus +
+ -
+ -
Vampirovibrio

+ - + +
Vampirovibrionaceae Genus

+ - + +
Deinococcota
Deinococcus


-


Meiothermus


-


Dependentiae
Babeliaceae Genus +

- + +
Desulfobacterota
Desulfobacterota Genus


- + +
Desulfobulbus


-

+
Elusimicrobiota
Lineage IIb Genus


-


Euryarchaeota
Methanobacteriaceae Genus


- +

Fibrobacterota
Fibrobacteraceae Genus


-
+
Firmicutes
Acidaminobacter


-
+ +
Anaerospora

+



Bacillus
+ - -


Brevibacillus
+




Clostridium sensu stricto 12


-
+
Clostridium sensu stricto 3

+ - + +
Enterococcus
+




Geothermomicrobium


-
+
Paraclostridium + + +
-

Staphylococcus

+



Streptococcus


-
+
Thermoactinomycetaceae Genus +
+



Vagococcus




+
ZOR0006

-
- -
Fusobacteriota
Cetobacterium + + -
-
-
Gemmatimonadota
Gemmatimonadaceae Genus
-
- + +
Halobacterota
Methanosarcina



+

MBNT15
MBNT15 Genus +
+ -
+
Myxococcota
Aetherobacter

+ - +

Anaeromyxobacter


-

+
Blfdi19 Genus


-


Haliangium

- -
+ +
P3OB 42


-
+
Pajaroellobacter +
+ -


Phaselicystis


-

+
Polyangia Genus


-


Sandaracinaceae Genus + + + - + +
mle1 27 Genus


-

+
NB1-j
NB1 j Genus
+ + - + +
Nitrospirota
Nitrospira
- + - + +
Patescibacteria
Saccharimonadales Genus + + + - +

TM7a



+

Planctomycetota
BD7 11 Genus


-

+
Blastopirellula +

- + +
CL500 3

+ - - +
Fimbriiglobus + - + - + +
Gemmata
+ + - + +
Gemmataceae Genus
-
- +

Isosphaeraceae Genus

+ - + + +
OM190 Genus


- + + +
Pir4 lineage
- + - + +
Pirellula + + + - -

Pirellulaceae Genus + - + - -

Planctomycetales Genus

+ - + +
Planctomycetes Genus

+ - + +
Planctomycetota Genus


-
+
Rhodopirellula


- + + -
SH PL14
+ + -


SM1A02

+ - + +
Schlesneria
-
- + +
Telmatocola

+ - + +
Tundrisphaera + - - -

-
vadinHA49 Genus


-
+
Proteobacteria
A0839 Genus +

- + +
AAP99


-
+
Acinetobacter -





Aeromonas -

+ +

Ahniella


-
+ +
Allorhizobium Neorhizobium Pararhizobium Rhizobium

-



Alphaproteobacteria Genus
+ + - + +
Alsobacter
+ + - + +
Aminobacter - -




Aquabacterium

-



Aquicella


- +

Azospira


- + +
Bauldia +
+ - + +
Beijerinckiaceae Genus
- + -
+ -
Blastomonas

-



Bosea - - - -


Burkholderia Caballeronia Paraburkholderia





+
Burkholderiales Genus


-


CCM19a Genus +

- + +
Caedibacter +
-
+

Candidatus Accumulibacter


-
+ -
Candidatus Alysiosphaera +
+ - + +
Candidatus Berkiella
- +
-

Candidatus Competibacter


-


Candidatus Megaira

+ - +

Candidatus Ovatusbacter
-
-


Caulobacter

-



Chitinibacter
+ +
+
-
Chitinilyticum

-
-

Chitinivorax


-
+
Comamonadaceae Genus

- - + +
Coxiella


-

+
Crenobacter
-




Curvibacter - -
-

+
Dechlorobacter


-
+
Dechloromonas


-
+
Defluviimonas
-
- -

Diplorickettsiaceae Genus


-


Elstera -



+
Fodinicurvataceae Genus


-
+
Ga0077536 Genus
-
-
+
Gammaproteobacteria Genus - - + -
+
Gemmobacter + + - -


Hirschia


-

+
Holosporaceae Genus - -

+

Hydrogenophaga





+
Hyphomicrobium
+ +



Ideonella


-
+ +
Inhella - -
-
+ +
Leeia - -
-
+
Legionella - -

-

Luteimonas -

-
+ +
Magnetospirillum


-
+
Mesorhizobium


-


Methyloligellaceae Genus + + + - + +
Mycoplana - - -



Neisseriaceae Genus

-



Nitrosomonadaceae Genus


-
+
Nitrosomonas
-
-
+
Nordella
-
- + +
Novosphingobium - -
-


Paucibacter + + - - -

Pedomicrobium + + +
-

Phenylobacterium


-

+
Phreatobacter


- -

Plesiomonas + +

- - -
Polymorphobacter
-
- + +
Porticoccus


-
+
Pseudomonadales Genus - - - - - -
Pseudomonas
+

+

Pseudorhodobacter
-
- + +
Pseudorhodoplanes +
+ - -

Reyranella
- + - + +
Rheinheimera -




+
Rhizobacter


- + + +
Rhizobiaceae Genus - -
-


Rhizobiales Incertae Sedis Genus


-
+
Rhizorhapis -

-

+
Rhodobacteraceae Genus


- + +
Rhodospirillales Genus


-

+
Rhodovarius


- + +
Rhodovastum
-
-


Rickettsiaceae Genus
+ +
+

Roseomonas + + + - - +
Rubrivivax


- -

SC I 84 Genus
-
- + +
SWB02


-

+
Shewanella - -



-
Shinella - -
-


Sphingopyxis





+
Sphingorhabdus -

-
+ +
Steroidobacter

+ -


Steroidobacteraceae Genus

+ - + +
Sulfurifustis





+
Sulfuritalea


-
+
Sutterellaceae Genus +
-
- -
TRA3 20 Genus





+
Tabrizicola

- - -
+
Undibacterium - -
-
+
Vibrio - -

+ + -
Xanthobacteraceae Genus - - + -
+
Zoogloea


-
+ -
alphaI cluster +
-
-

Spirochaetota
Turneriella - -
-
+
Verrucomicrobiota
Candidatus Protochlamydia


- + +
Chlamydiales Genus

+ -
+ -
Chthoniobacter
-
- - + -
DEV008


-

+
Luteolibacter


-
+
Neochlamydia

+ - -

Oikopleura

+ - + +
Opitutaceae Genus


-
+
Parachlamydiaceae Genus

-



Pedosphaeraceae Genus
-
- + +
SH3 11

- - -

Simkania
-




Simkaniaceae Genus

+



Verrucomicrobiaceae Genus - -
-

+
Verrucomicrobiae Genus

-



WPS-2
WPS 2 Genus -




+

Session Info

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Session Info

## R version 4.3.3 (2024-02-29)
## Platform: x86_64-apple-darwin20 (64-bit)
## Running under: macOS 15.3
## 
## Matrix products: default
## BLAS:   /System/Library/Frameworks/Accelerate.framework/Versions/A/Frameworks/vecLib.framework/Versions/A/libBLAS.dylib 
## LAPACK: /Library/Frameworks/R.framework/Versions/4.3-x86_64/Resources/lib/libRlapack.dylib;  LAPACK version 3.11.0
## 
## locale:
## [1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8
## 
## time zone: America/Los_Angeles
## tzcode source: internal
## 
## attached base packages:
## [1] grid      parallel  stats     graphics  grDevices utils     datasets 
## [8] methods   base     
## 
## other attached packages:
##  [1] lubridate_1.9.3       dplyr_1.1.4           readr_2.1.5          
##  [4] tidyr_1.3.1           tibble_3.2.1          tidyverse_2.0.0      
##  [7] multcomp_1.4-25       TH.data_1.1-2         MASS_7.3-60.0.1      
## [10] survival_3.6-4        mvtnorm_1.2-5         emmeans_1.10.1       
## [13] lme4_1.1-35.3         Matrix_1.6-5          car_3.1-2            
## [16] carData_3.0-5         broom_1.0.6           microViz_0.12.1      
## [19] picante_1.8.2         nlme_3.1-164          ape_5.8              
## [22] microbiome_1.24.0     phyloseqCompanion_1.1 vegan_2.6-4          
## [25] lattice_0.22-6        permute_0.9-7         stringr_1.5.1        
## [28] rlang_1.1.4           magrittr_2.0.3        GUniFrac_1.8         
## [31] doParallel_1.0.17     iterators_1.0.14      foreach_1.5.2        
## [34] data.table_1.15.4     phyloseq_1.46.0       Maaslin2_1.16.0      
## [37] ggnewscale_0.4.10     ComplexHeatmap_2.18.0 RColorBrewer_1.1-3   
## [40] gt_0.10.1             ggpubr_0.6.0          ggrepel_0.9.5        
## [43] ggExtra_0.10.1        ggbeeswarm_0.7.2      ggplot2_3.5.1        
## [46] furrr_0.3.1           future_1.33.2         purrr_1.0.2          
## [49] scales_1.3.0          rcompanion_2.4.35     knitr_1.45           
## [52] forcats_1.0.0        
## 
## loaded via a namespace (and not attached):
##   [1] fs_1.6.4                matrixStats_1.3.0       bitops_1.0-7           
##   [4] httr_1.4.7              insight_0.19.10         tools_4.3.3            
##   [7] backports_1.5.0         utf8_1.2.4              R6_2.5.1               
##  [10] mgcv_1.9-1              nortest_1.0-4           rhdf5filters_1.14.1    
##  [13] GetoptLong_1.0.5        withr_3.0.0             cli_3.6.2              
##  [16] Biobase_2.62.0          textshaping_0.3.7       logging_0.10-108       
##  [19] biglm_0.9-2.1           sandwich_3.1-0          labeling_0.4.3         
##  [22] sass_0.4.9              robustbase_0.99-2       pbapply_1.7-2          
##  [25] proxy_0.4-27            systemfonts_1.0.6       parallelly_1.37.1      
##  [28] readxl_1.4.3            rstudioapi_0.16.0       generics_0.1.3         
##  [31] shape_1.4.6.1           vroom_1.6.5             zip_2.3.1              
##  [34] inline_0.3.19           biomformat_1.30.0       fansi_1.0.6            
##  [37] DescTools_0.99.54       S4Vectors_0.40.2        abind_1.4-5            
##  [40] juicyjuice_0.1.0        lifecycle_1.0.4         yaml_2.3.8             
##  [43] rhdf5_2.46.1            Rtsne_0.17              promises_1.3.0         
##  [46] crayon_1.5.2            miniUI_0.1.1.1          cowplot_1.1.3          
##  [49] chromote_0.2.0          pillar_1.9.0            optparse_1.7.5         
##  [52] statip_0.2.3            rjson_0.2.21            boot_1.3-30            
##  [55] gld_2.6.6               estimability_1.5        codetools_0.2-20       
##  [58] glue_1.7.0              V8_4.4.2                vctrs_0.6.5            
##  [61] png_0.1-8               cellranger_1.1.0        gtable_0.3.5           
##  [64] datawizard_0.10.0       openxlsx_4.2.8          xfun_0.43              
##  [67] mime_0.12               libcoin_1.0-10          coda_0.19-4.1          
##  [70] rsconnect_1.3.4         pcaPP_2.0-4             modeest_2.4.0          
##  [73] timeDate_4032.109       pheatmap_1.0.12         statmod_1.5.0          
##  [76] bit64_4.0.5             modelbased_0.8.7        GenomeInfoDb_1.38.8    
##  [79] fBasics_4032.96         vipor_0.4.7             rpart_4.1.23           
##  [82] colorspace_2.1-0        BiocGenerics_0.48.1     DBI_1.2.3              
##  [85] ade4_1.7-22             processx_3.8.4          Exact_3.2              
##  [88] tidyselect_1.2.1        timeSeries_4032.109     curl_5.2.1             
##  [91] bit_4.0.5               compiler_4.3.3          expm_0.999-9           
##  [94] xml2_1.3.6              DEoptimR_1.1-3          lmtest_0.9-40          
##  [97] spatial_7.3-17          multcompView_0.1-10     digest_0.6.35          
## [100] minqa_1.2.6             rmarkdown_2.26          XVector_0.42.0         
## [103] htmltools_0.5.8.1       pkgconfig_2.0.3         openxlsx2_1.14         
## [106] stabledist_0.7-1        fastmap_1.2.0           GlobalOptions_0.1.2    
## [109] shiny_1.8.1.1           farver_2.1.2            zoo_1.8-12             
## [112] jsonlite_1.8.8          RCurl_1.98-1.14         modeltools_0.2-23      
## [115] GenomeInfoDbData_1.2.11 Rhdf5lib_1.24.2         munsell_0.5.1          
## [118] Rcpp_1.0.12             stringi_1.8.4           rootSolve_1.8.2.4      
## [121] stable_1.1.6            zlibbioc_1.48.2         plyr_1.8.9             
## [124] listenv_0.9.1           lmom_3.0                Biostrings_2.70.3      
## [127] splines_4.3.3           hash_2.2.6.3            multtest_2.58.0        
## [130] hms_1.1.3               circlize_0.4.16         ps_1.7.6               
## [133] igraph_2.0.3            ggsignif_0.6.4          reshape2_1.4.4         
## [136] stats4_4.3.3            rmutil_1.1.10           evaluate_0.23          
## [139] BiocManager_1.30.23     nloptr_2.0.3            tzdb_0.4.0             
## [142] httpuv_1.6.15           webshot2_0.1.1          getopt_1.20.4          
## [145] clue_0.3-65             coin_1.4-3              xtable_1.8-4           
## [148] e1071_1.7-14            rstatix_0.7.2           later_1.3.2            
## [151] class_7.3-22            ragg_1.3.0              report_0.5.8           
## [154] websocket_1.4.1         beeswarm_0.4.0          IRanges_2.36.0         
## [157] writexl_1.5.2           cluster_2.1.6           timechange_0.3.0       
## [160] globals_0.16.3